Fluorescent nanoparticles to improve resections of microscopic pancreatic tumors

摘要 胰腺癌的一个主要挑战是预防患者在“治愈意图”后的肿瘤复发 切除术（5年生存率21%）。据信，在一部分患者中， 肿瘤复发是小的亚厘米和亚毫米的疾病，是看不见的，因此，未切除的时候， 手术为了解决这个问题，研究人员正在开发新的辅助引导细胞减灭术 （FGCS）技术，旨在使外科医生能够在术中检测和切除亚厘米和亚毫米 肿瘤的这些技术施用荧光探针-例如荧光缀合的抗体、小的荧光探针、荧光探针、荧光标记的抗体或荧光标记的抗体。 分子、肽、量子点或纳米颗粒-其定位于肿瘤，从而有助于识别 肿瘤组织肿瘤定位通常通过两种广泛分类的靶向策略之一来实现： 通过增强的渗透性和保留（EPR）效应的"被动靶向";或者，通过增强的渗透性和保留（EPR）效应的"主动靶向"。 将靶向部分或抗体掺入探针中。然而，这些技术都不能 识别亚厘米和亚毫米疾病。该提案开发了一种新颖的、获得专利的荧光探针，即"高度- 荧光罗丹明标记的可膨胀纳米颗粒"（HFR-eNP），通过独特的材料靶向肿瘤- 基于定位战略。这种机制利用了纳米颗粒聚合物的材料功能性和纳米颗粒聚合物的材料功能性。 (e.g.,肿胀）和肿瘤的基本病理生理学特性（例如，代谢率增加）， 实现肿瘤特异性定位，准确率> 95%。我们假设，通过使用HFR-eNPs来引导 对播散性亚cm和亚mm肿瘤进行细胞减灭术，我们将显著提高总体生存率 与无引导切除相比。初步数据表明：1）HFR-eNPs具有5至10倍的增加， 与等同浓度的游离罗丹明相比， 照射而没有这种荧光的显著损失; 2）在临床上大规模生产HFR-eNPs（即， 1升批次）规模; 3）HFR-eNPs在体外和体内的无毒性; 4）对HFR-1的敏感性和特异性定位; 体内亚cm和亚mm胰腺肿瘤;和，5）概念验证HFR-eNP引导的细胞减灭术 以在体内去除大的（> 1cm）、亚cm和亚mm肿瘤。关于COM的两个关键的Go/No-Go决定- 该技术的商业化在本提案中得到了解决。首先，为了在临床上使用， 必须开发不改变HFR-eNP聚合物或淬灭罗丹明荧光的方案。 并且，灭菌后，必须确认颗粒的体内功能性/肿瘤定位。第二、 - 通过使用HFR-eNPs引导胰腺肿瘤（特别是亚cm）切除所提供的益处， 必须通过体内细胞减灭手术模型来定量。因此，这一目标 目的1）通过γ射线照射对HFR-eNPs进行灭菌，并确认灭菌后的肿瘤定位; 目的2）确定HFR-eNP引导的手术切除对生存率的改善。

ABSTRACT A primary challenge in pancreatic cancer is preventing tumor recurrence in patients following a “curative intent” resection procedure (5-year survival rate 21%). It is believed that, in a subset of patients, the primary cause of tumor recurrence is small sub-cm and sub-mm disease that is unseen and, therefore, un-resected at the time of surgery. To address this problem, researchers are developing new Fluorescently-Guided Cytoreductive Surgery (FGCS) techniques with the goal of enabling surgeons to intraoperatively detect and resect sub-cm and sub-mm tumors. These techniques administer a fluorescent probe—such as a fluorescently-conjugated antibody, small molecule, peptide, quantum dot or nanoparticle—that localizes to tumors thereby facilitating identification of tumor tissue. Localization to the tumor is generally achieved via one of two broadly classified targeting strategies: “Passive Targeting” via the enhanced-permeability and retention (EPR) effect; or, “Active Targeting” through the incorporation of a targeting moiety or antibody into the probe. However, none of these technologies is able to identify sub-cm and sub-mm disease. This proposal develops a novel, patented, fluorescent probe, the “highly- fluorescent rhodamine-labeled expansile nanoparticle” (HFR-eNP), that targets tumors via a unique Materials- Based Targeting strategy. This mechanism leverages both the material functionality of the nanoparticle polymer (e.g., swelling) and fundamental pathophysiological properties of tumors (e.g., increased metabolic rate) to achieve tumor-specific localization with >95% accuracy. We hypothesize that by using HFR-eNPs to guide cytoreductive surgery of disseminated sub-cm and sub-mm tumors, we will significantly improve overall survival compared to unguided resections. Preliminary data demonstrate: 1) HFR-eNPs possess 5- to 10-fold increased fluorescence compared to equivalent concentrations of free rhodamine and can be sterilized with gamma irradiation without significant loss of this fluorescence; 2) large-scale production of HFR-eNPs on a clinical (i.e., 1 liter batch) scale; 3) non-toxicity of the HFR-eNPs in vitro and in vivo; 4) sensitive and specific localization to sub-cm and sub-mm pancreatic tumors in vivo; and, 5) proof-of-concept HFR-eNP-guided cytoreductive surgery to remove large (>1 cm), sub-cm and sub-mm tumors in vivo. Two key Go/No-Go decisions regarding the com- mercialization of this technology are addressed in this proposal. First, in order to be used clinically, a sterilization protocol must be developed that does not alter the HFR-eNP polymer or quench the rhodamine fluorescence. And, following sterilization, the in vivo functionality/tumor localization of the particles must be confirmed. Second, the benefit afforded by using HFR-eNPs to guide the resection of pancreatic tumors, in particular sub-cm and sub-mm tumors, must be quantified through an in vivo cytoreductive surgery model. Thus, the aims of this proposal are: Aim 1) Sterilize HFR-eNPs via Gamma irradiation and confirm tumor localization post-sterilization; Aim 2) Determine the improvement to survival afforded by HFR-eNP-guided surgical resections.