Cancer Margin Detection using Infrared Spectroscopy and Plasmon Resonance

描述（由申请人提供）：使用红外光谱和等离子体共振的癌边缘检测摘要虽然癌组织的手术切除是对多种形式的人类癌症的单一最有效的治疗，但是外科手术的有效性受到准确识别肿瘤边缘的困难的限制。因此，使用离体免疫组织化学（IHC）开发癌症特异性分子表面探针是高度期望的。在拟定的研究中，将使用具有表面等离子体网格增强的红外（IR）光谱来评估正常组织与腺癌组织。这项技术提供了生物组织的分子特征。此外，我们将利用标记的IR报告分子系统，该系统由合成连接到抗肿瘤相关糖蛋白-72（抗TAG-72）的氘代聚乙二醇（dPEG）分子组成，所述抗TAG-72选择性地与腺癌组织共定位。这解决了在手术期间离体区分正常组织和癌组织的关键需求。IR方法，特别是使用具有高度可再现IR光谱的表面等离子体增强网材料的IR方法，具有克服当前耗时且劳动密集型组织学程序的限制的潜力。肿瘤边缘的特异性分子表征将比目前依赖于肿瘤活检材料的目视检查的方法更精确和可靠。先进的IR方法可以对切除的肿瘤组织进行全面分析，提高手术室组织学分析的质量，并指导癌组织的完全手术切除。该方案的方法是构建dPEG标记的腺癌特异性抗TAG-72单克隆抗体（MAbs），并评估这些报告基因用于识别肿瘤边缘的用途。IR光谱和标记的抗TAG MAb的组合能够减少评估时间并向外科医生提供及时反馈。它还可以减少不必要的破坏或切除正常组织，改善患者的预后。抗TAG单克隆抗体已被证明对结肠癌、乳腺癌和妇科癌组织具有高度选择性，使得该方法适用于一系列医学上重要的人类癌症。用于本拟议研究的结肠、乳腺和盆腔/生殖器组织的人体组织样本将由Martin、Povoski和O 'Malley博士组成的外科肿瘤学团队提供。病理学团队希区柯克博士和艾伦博士将领导组织学和分析方法的开发，包括使用报告构建体进行组织染色的程序。此外，Coe将领导表面等离子体激元IR增强方法策略。团队成员（PI团队和手术团队）在过去几年中在MAb开发方面建立了工作关系，目前的提案汇集了IR和增强技术方面的额外专业知识，已证明具有协同作用。 公共卫生相关性：表面等离子体增强红外显微光谱将与肿瘤特异性抗体结合使用，以开发一种快速，灵敏，准确的方法来分析切除的癌组织的分子特征。该方法将构建由氘代聚乙二醇（PEG）标记的分子与腺癌特异性抗肿瘤相关糖蛋白-72（anti-TAG-72）单克隆抗体偶联组成的红外报告系统，用于肿瘤边缘的组织学评估。

DESCRIPTION (provided by applicant): Cancer Margin Detection using Infrared Spectroscopy and Plasmon Resonance Abstract While surgical resection of cancerous tissue is the single most effective treatment for many forms of human cancer, the effectiveness of surgical procedures is limited by the difficulty of accurately recognizing tumor margins. Thus, the development of cancer specific molecular surface probes using ex vivo immunohistochemistry (IHC) is highly desirable. In the proposed studies, infrared (IR) spectroscopy with surface plasmon mesh enhancement will be used to assess normal versus adenocarcinoma tissues. This technology provides molecular signatures of biological tissue. Moreover, we will utilize labeled IR reporter molecular systems that consist of deuterated polyethylene glycol (dPEG) molecules synthetically attached to anti-tumor-associated-glycoprotein- 72 (anti-TAG-72), which selectively co-locates with adenocarcinoma tissue. This addresses the critical need for ex vivo differentiation between normal and cancerous tissue during surgery. IR methods, specifically with surface plasmon enhancement mesh materials that have highly reproducible IR absorbances, have the potential to overcome the limitations of current time consuming and labor intensive histological procedures. The specific molecular characterization of tumor margins will be more precise and reliable than current methods which depend on visual inspection of tumor biopsy material. Advanced IR methods can enable thorough analysis of resected tumor tissue, increase the quality of operating room histological analyses, and guide complete surgical removal of cancerous tissue. The approach of this proposal is to construct dPEG labeled adenocarcinomo-specific anti-TAG-72 monoclonal antibodies (MAbs) and assess the use of these reporters to identify tumor margins. The combination of IR spectroscopy and labeled anti-TAG MAbs has the ability to reduce assessment time and to supply timely feedback to the surgeon. It can also reduce the needless destruction or removal of normal tissues, improving patient outcomes. Anti-TAG MAbs have been shown to be highly selective for colon, breast, and gynecological cancer tissues making this methodology applicable to a range of medically important human cancers. Human tissue samples from colon, breast and pelvic/genital tissues for this proposed research will be provided by a surgical oncology team comprised of Drs. Martin, Povoski, and O'Malley. The pathology team, Drs. Hitchcock and Allen, will lead the development of histological and analytical methods including procedures for tissue staining with reporter constructs. Additionally, Coe will lead the surface plasmon IR enhancement methodology strategies. Team members (PI team and surgical team) have an established working relationship in MAb development in past years and this current proposal brings together additional expertise in IR and enhancement technology that has shown to be synergistic. PUBLIC HEALTH RELEVANCE: Surface plasmon enhanced infrared microspectroscopy will be used in combination with tumor specific antibodies to develop a rapid, sensitive, and accurate method to analyze the molecular characteristics of resected cancerous tissues. The approach will be to construct an infrared reporter system consisting of deuterated polyethylene glycol (PEG) labeled molecules conjugated to adenocarcinoma-specific anti-tumor-associated-glycoprotein-72 (anti-TAG-72) monoclonal antibodies for histological assessment of tumor margins.