Novel probes for near-infrared fluorescence imaging-guided oncologic surgery

用于近红外荧光成像引导肿瘤手术的新型探针

• 批准号: 9025567
• 负责人: VICTOR KRASNYKH
• 金额: $ 8万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-04-01 至 2017-09-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9025567
• 关键词: Affinity; Animal Cancer Model; Ankyrin Repeat; Antibodies; Bacteria; Blood; Blood Circulation; Cancer Patient; Cancerous; Clinical; Detection; Development; Diagnostic; Diagnostic Imaging; Disease; Engineering; Epidermal Growth Factor Receptor; Excision; Eye; Feasibility Studies; Fluorescence; Fluoroscopy; Generations; Goals; Health; Human; Image; Imagery; Imaging technology; In Vitro; Lesion; Life; Light; Literature; Location; Malignant Neoplasms; Malignant neoplasm of ovary; Microscopic; Molecular; Molecular Target; Morbidity - disease rate; Near-Infrared Fluorescence Imaging Probe; Near-infrared optical imaging; Normal tissue morphology; Operative Surgical Procedures; Outcome; Palpation; Peptides; Pilot Projects; Postoperative Period; Property; Proteins; Public Health; Publishing; Radioisotopes; Research; Safety; Signal Transduction; Site; Specificity; Structure; Surgeon; Surgical Oncology; Technology; Testing; Therapeutic; Time; Tissues; Translating; Tumor Markers; Ultrasonography; Visual; base; clinical practice; design; fluorophore; image guided; imaging probe; improved; instrumentation; mouse model; novel; operation; optical imaging; prevent; programs; small molecule; soft tissue; tumor

 DESCRIPTION (provided by applicant): The outcomes of oncologic surgery critically depend on complete removal of the tumor mass, which prevents tumor regrowth and disease reoccurrence. This is a challenging task, especially in cases of invasive and disseminated disease in which tumor margins are poorly defined and microscopic tumors are often indistinguishable from surrounding normal tissues. Preoperative diagnostic imaging provides a wealth of information on the tumors' location and size. However, during surgery, surgeons identify tumors and tumor margins largely by visual inspection of the surgical field and by palpation; as a result, tumors often remain undetected, and tumor margins are determined incorrectly. On the other hand, excessive resection of tumor margins often leads to severe postoperative morbidity. These problems may be solved using real- time intraoperative visualization of tumors enabled by near-infrared fluorescence (NIRF), which would give a surgeon the ability to locate and remove the tumors that cannot be seen by the naked eye and enable more precise determination of tumor margins. The objective of this pilot project is to demonstrate the feasibility of sensitive and accurate intraoperative tumor visualization by developing and testing in an animal model of cancer a new generation of tumor-targeted NIRF probes. Our approach is based on the hypothesis that such advanced probes can be developed using a novel class of rationally engineered molecules-designed ankyrin repeat proteins (DARPins)-as carriers of NIR fluorophores. Owing to their small size, high affinity and specificity for tumor biomarkers, DARPins rapidly and efficiently accumulate in tumors and clear from nontarget tissues. DARPins are easy to produce in bacteria and purify; they can be easily modified genetically to enable precise, site-specific conjugation with fluorophores. DARPins' highly stable core structure results in prolonged stability in human blood. These properties make DARPins an excellent alternative to antibodies (Ab), Ab derivatives, peptides, and small molecules, which are traditionally used for imaging probe development. We will test our hypothesis and achieve our objective by pursuing two specific aims: Specific Aim 1. Develop and characterize in vitro DARPin-based NIRF probes that target human epidermal growth factor receptor 2 (Her2). Specific Aim 2. Demonstrate the feasibility of intraoperative visualization of disseminated tumors by using DARPin-derived NIRF probes in a mouse model of ovarian cancer. The outcomes of this feasibility project will have a significant positive impact on cancer management by making surgical resection of tumors more efficient and reducing post-surgical morbidity, thereby prolonging and saving human life.

 描述（由申请人提供）：肿瘤手术的结果关键取决于肿瘤块的完全切除，这可以防止肿瘤再生和疾病复发。这是一项具有挑战性的任务，特别是在侵袭性和播散性疾病的情况下，肿瘤边缘界限不清，微观肿瘤通常与周围正常组织难以区分。术前诊断成像提供了有关肿瘤位置和大小的丰富信息。然而，在手术过程中，外科医生主要通过目视检查手术区域和触诊来识别肿瘤和肿瘤边缘；结果，肿瘤常常未被发现，并且肿瘤边缘的确定不正确。另一方面，肿瘤边缘的过度切除往往会导致严重的术后并发症。这些问题可以通过近红外荧光（NIRF）实现肿瘤的实时术中可视化来解决，这将使外科医生能够定位和切除肉眼无法看到的肿瘤，并能够更精确地确定肿瘤边缘。该试点项目的目标是通过在癌症动物模型中开发和测试新一代肿瘤靶向 NIRF 探针，证明灵敏且准确的术中肿瘤可视化的可行性。我们的方法基于这样的假设：可以使用一类新型的合理工程分子——设计的锚蛋白重复蛋白（DARPins）——作为近红外荧光团的载体来开发这种先进的探针。由于 DARPins 尺寸小、对肿瘤生物标志物具有高亲和力和特异性，DARPins 可以快速有效地在肿瘤中积累，并从非靶组织中清除。 DARPins很容易在细菌中产生并纯化；它们可以很容易地进行基因修饰，以实现与荧光团的精确、位点特异性结合。 DARPins 高度稳定的核心结构可在人体血液中实现长期稳定性。这些特性使 DARPins 成为传统上用于成像探针开发的抗体 (Ab)、Ab 衍生物、肽和小分子的绝佳替代品。 我们将通过追求两个具体目标来检验我们的假设并实现我们的目标： 具体目标 1. 开发并表征体外基于 DARPin 的 NIRF 探针，该探针针对人表皮生长因子受体 2 (Her2)。具体目标 2. 证明在卵巢癌小鼠模型中使用 DARPin 衍生的 NIRF 探针实现术中播散性肿瘤可视化的可行性。该可行性项目的结果将对癌症治疗产生显着的积极影响，使肿瘤的手术切除更加有效，降低术后发病率，从而延长和挽救人类生命。