Optical imaging of a CD44-based oral-rinse with fluorescent detection to visualize oral cancer

基于 CD44 的口腔冲洗液的光学成像与荧光检测以可视化口腔癌

• 批准号: 9047943
• 负责人: Elizabeth J Franzmann
• 金额: $ 19.15万
• 依托单位: VIGILANT BIOSCIENCES, INC.
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-02-15 至 2017-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9047943
• 关键词: Abnormal Cell; Address; Aerodigestive Tract; Affect; Alcohols; Antibodies; Area; Bedside Testings; Benign; Binding; Biopsy; CD44 gene; Cell Count; Cell Line; Cell surface; Cells; Chronic; Clinical; Confocal Microscopy; Cost Savings; Data; Death Rate; Detection; Diagnosis; Diagnostic Neoplasm Staging; Disease; Early Diagnosis; Early Intervention; Engineering; Epithelium; Excision; Eye; Fluorescein-5-isothiocyanate; Fluorescence; Fluorescent Probes; Gold; Health; Health Care Costs; Histology; Human; Image; Imagery; Imaging Device; Incubated; Individual; Label; Lateral; Lesion; Light; Location; Malignant Neoplasms; Measures; Medical; Membrane; Microspheres; Morbidity - disease rate; Operative Surgical Procedures; Oral; Oral cavity; Oropharyngeal; Outcome; Patient risk; Patients; Proteins; Radiosurgery; Research; Risk; Screening for Oral Cancer; Specimen; Staging; Structure; Survival Rate; Symptoms; System; Testing; Tissues; Tobacco; Tonsillitis; Treatment Cost; Treatment Protocols; Ultraviolet Rays; Visual; antibody conjugate; base; cancer diagnosis; carcinogenesis; chemotherapy; fluorescence imaging; imaging agent; improved; in vitro Model; lymph nodes; malignant mouth neoplasm; mortality; mouth squamous cell carcinoma; optical imaging; overexpression; programs; tumor

DESCRIPTION (provided by applicant): Oral squamous cell carcinomas (including those of the mouth and oropharynx) comprise more than 90% of all cancers affecting these tissues. Of all the major cancers, oral cancer has one of the worst five-year survival rates of 62.7%. Historically, the death rate associated with this cancer is particularly high because it is discovered late (stage III or IV), often when it has spread to the lymph nodes or deeper structures. Late stage cancers have high mortality, are expensive to treat and have high morbidity. The current "gold standard" for oral cancer diagnosis is a physical and visual examination followed by biopsy. Unfortunately, due to delays in diagnosis, most cases are diagnosed in late stage with a survival rate around 20-40% even after aggressive treatment regimens including combinations of radiation, surgery and chemotherapy. However, if diagnosed early (stage I/II), the five-year survival-rate can be as high as 80-90%, representing a significant healthcare cost savings. Thus, there is a pressing need for an improved oral cancer early detection test which will save lives, improve clinical outcome and reduce treatment costs. We have developed a unique, easy to use, oral cancer specific, rapid lateral flow point-of-care (POC) test based on the tumor initiating marker CD44 and total protein that accurately assesses patient's risk for oral cancer. The OncAlert lateral flow test strip identifies those individuals a most risk of harboring early or developing oral cancer from the 85 million tobacco and alcohol users. However, often these early lesions can be invisible to the naked eye, so the test strip alone will not indicate where the abnormal cells are in cases of very early disease. To answer this pressing medical need, in this application, we will develop an imaging device that will involve a simple rinse and spit solution with an antiCD44 probe bound to non-absorbable microspheres that will bind to cells expressing abnormally high levels of CD44. The bound probe can then be visualized by shining a light of specific wavelength so that very early lesions can be identified. If successful, this is expected to enable early detection and intervention of orl cancer even be- fore visual symptoms are revealed, thus addressing a crucial clinically unmet need. We will carry out the fol- lowing specific aims: in Specific Aim #1 we will test an in-vitro model for detecting CD44 elevated cells using fluorescent probes on membranes. We will determine whether this potential imaging agent recognizes CD44 on the surface of cells. To do this we will culture well-characterized cell lines that have been engineered to overexpress or underexpress CD44. In Specific Aim #2 we will test the system in tumor explants from humans. We will determine how well the conjugate identifies carcinogenesis in humans. We will use human up- per aero-digestive tract specimens ex vivo, immediately following surgical resection. We will analyze 10 oral cancer tissues with surrounding normal margin and 10 patient tissues that were excised for benign conditions (chronic tonsillitis) using the anti-CD44 fluorescently labeled beads.

描述（由申请人提供）：口腔鳞状细胞癌（包括口腔和口咽部的鳞状细胞癌）占影响这些组织的所有癌症的 90% 以上。在所有主要癌症中，口腔癌的五年生存率最差，为 62.7%。从历史上看，与这种癌症相关的死亡率特别高，因为它是在晚期（III期或IV期）发现的，通常是在它已经扩散到淋巴结或更深的结构时。晚期癌症死亡率高、治疗费用高且发病率高。目前口腔癌诊断的“金标准”是进行身体检查和目视检查，然后进行活检。不幸的是，由于诊断延误，大多数病例在晚期才被诊断出来，即使经过积极的治疗方案（包括放射、手术和化疗的组合），生存率仍约为 20-40%。然而，如果早期诊断（I/II 期），五年生存率可高达 80-90%，这意味着可节省大量医疗费用。因此，迫切需要改进的口腔癌早期检测测试，以挽救生命、改善临床结果并降低治疗成本。我们开发了一种独特、易于使用、针对口腔癌的快速侧流即时护理 (POC) 测试，该测试基于肿瘤起始标记物 CD44 和总蛋白，可准确评估患者患口腔癌的风险。 OncAlert 侧流层析测试条从 8500 万烟草和酒精使用者中识别出那些最有可能罹患早期或发展为口腔癌的人。然而，这些早期病变通常是肉眼看不见的，因此在非常早期的疾病中，仅靠试纸条并不能指示异常细胞的位置。为了满足这一紧迫的医疗需求，在本应用中，我们将开发一种成像设备，该设备涉及简单的冲洗和吐出溶液，其中包含与不可吸收微球结合的抗CD44探针，该微球将与表达异常高水平CD44的细胞结合。然后可以通过照射特定波长的光来观察结合的探针，从而可以识别出非常早期的病变。如果成功，预计甚至可以在视觉症状显现之前早期发现和干预口腔癌，从而解决临床上未满足的关键需求。我们将实现以下具体目标：在具体目标#1中，我们将测试体外模型，使用膜上的荧光探针检测CD44升高的细胞。我们将确定这种潜在的成像剂是否识别细胞表面的 CD44。为此，我们将培养经过精心设计的细胞系，这些细胞系被设计为过表达或低表达 CD44。在具体目标#2 中，我们将在人类肿瘤外植体中测试该系统。我们将确定该缀合物识别人类致癌作用的效果。我们将在手术切除后立即使用离体的人类上呼吸消化道标本。我们将使用抗 CD44 荧光标记珠分析 10 个周围正常边缘的口腔癌组织和 10 个因良性疾病（慢性扁桃体炎）而切除的患者组织。