Imaging Nonlinear Absorption of Biomarkers for Improved Detection of Melanoma

生物标志物的非线性吸收成像可改善黑色素瘤的检测

基本信息

批准号：
7942928
负责人：
Warren S Warren
金额：
$ 49.85万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-30 至 2012-03-29
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7942928
关键词：
Address Adjuvant Therapy American Animal Model Area Basic Science Benign Biological Markers Biomedical Engineering Blood Blood flow Cell Culture Techniques Characteristics Chemistry Clinical Clinical Research Dermatologist Dermoscopy Detection Development Diagnosis Diagnostic Evaluation Event Excision Face Fluorescence Frequencies Funding Goals Grant Hair Health Health Care Costs Hemoglobin Histology Histopathology Human Image Imaging technology Langerhans cell Laser Scanning Confocal Microscopy Lasers Lead Lesion Life Life Insurance Light Malignant - descriptor Malignant Neoplasms Malpractice Measures Melanins Melanosomes Metabolism Methods Microscope Microscopic Microscopy Mole the mammal Molecular Molecular Profiling Molecular Target Neoplasms Nude Mice Optics Oxyhemoglobin Pathologist Patients Physics Physiologic pulse Pigmentation physiologic function Pigments Probability Procedures Process Public Health Radiology Specialty Relative (related person)Reporting Research Resolution Sampling Science Sensitivity and Specificity Sentinel Lymph Node Biopsy Shapes Skin Skin Cancer Skin Transplantation Skin graft Societies Specificity Staging Staining method Stains Structure Surface System Techniques Technology Testing Tissue Sample Tissues Training United States Validation Visual Work absorption clinical application deoxyhemoglobin design eumelanin human tissue improved in vivo insight interest light scattering malignant breast neoplasm melanocyte melanoma molecular imaging molecular marker mortality mouse model multi-photon novel penis foreskin pheomelanin professor public health relevance research study skin lesion tissue oxygenation

项目摘要

DESCRIPTION (provided by applicant): This application addresses broad Challenge Area Biomarker Discovery and Validation (03) and specific Challenge Topic Imaging Biomarkers (03-AR-104). We desire to detect important biomarkers associated with the onset of skin cancers, with a particular emphasis on melanoma. Specifically, we will apply a novel imaging technology (transient absorption microscopy) to image melanins and hemoglobins in developing skin lesions, in both fixed and live skin. The goals are to noninvasively detect melanomas developing in their earliest stages and to reduce false positives and false negatives in both dermoscopy and histopathology. Conventional melanoma detection and diagnosis is a two-tier process that begins with visual or dermoscopic inspection of suspicious moles and ends with the removal of suspected tissue that is examined in a microscope to confirm diagnosis. However, this approach faces two fundamental challenges. The first is the difficulty in visually detecting the differences between melanoma and benign moles; most melanomas are highly pigmented, and even using a dermoscope, doctors cannot see far beneath the surface. This problem is largely addressed by excision, H&E staining, and examination by a pathologist, although patients generally present far too many moles to excise and test in such an invasive manner. The second challenge is that false negatives and false positives from histopathology remain a serious problem even with trained observers. False negatives delay treatment and increase mortality; in fact misdiagnosis of melanoma is the second most common reason for cancer malpractice claims in the United States (after breast cancer). False positives drive up the cost of healthcare with unnecessary, expensive, and invasive procedures (including sentinel lymph node biopsy and systemic adjuvant therapy) and may make it impossible for the patient to obtain health or life insurance. Thus, more accurate detection and diagnosis could have a very large impact on patient survival and health care cost reduction. Existing microscopy methods are not well suited to deal with these challenges. Reflectance confocal scanning laser microscopy (rCSLM) has been commercialized for this application, but the contrast (scattering) suffers from a lack of specificity as it does not target a specific biomarker. Conventional multiphoton microscopy can image deep into tissue with microscopic resolution, but pigmented lesions present a horrible target because the generated light is reabsorbed (and in any event the fluorescence from melanin is extremely weak). Here we use nonlinear transient absorption microscopy, which does not retains the resolution advantage of multiphoton microscopy but does require the sample to generate light at a new wavelength. Such methods have been around for decades, but recent work in the PI's lab has exploited advanced femtosecond pulse shaping and pulse train modulation methods to dramatically increase the sensitivity- thus making it feasible to image tissue with modest powers. He and his research group have migrated this technology from basic science to clinical applications. The targeted biomarkers are significant for melanoma diagnosis. There is good evidence that the local ratio between pheomelanin and eumelanin contents are altered in developing melanoma; here we dramatically improve on previous work by measuring this distribution microscopically (in melanosomes and melanocytes), at depth in tissue, and noninvasively. There is also evidence that microvascularity and oxygenation (oxy- and deoxyhemoglobin, which we can also image noninvasively and at depth) correlates with metastatic potential. Overall, the research team in this project includes chemists, laser technologists, pathologists and dermatologists in order to truly gather the expertise needed to make a clinical impact. For example, the PI is a pioneer in molecular imaging and ultrafast laser physics (and Chair-Elect of the Division of Laser Science of the American Physical Society), but also a professor in Chemistry, Radiology and Biomedical Engineering. In the first aim, we focus on analyzing de-identified excised moles with our new methods and with the best conventional methods. As this work evolves, and we determine the molecular signatures which best correlate with cancer development, we will progress to freshly excised skin studies, in order to validate safe power limits (even though we currently use less laser power than existing commercial systems). Finally, we progress to a live animal model (human skin with induced lesions grafted on nude mice), where we will also measure blood flow and local tissue oxygenation as likely markers of aggressive metabolism. By the end of the grant period, we will be poised to provide spectacular and novel insight into the development of melanoma in vivo, and be ready for human clinical work. PUBLIC HEALTH RELEVANCE: We propose a novel imaging technology which can image specific cancer biomarkers in developing skin lesions, to noninvasively detect early melanomas without excision and to reduce false positives and false negatives in histopathology. Reducing false negatives would reduce cancer fatalities; reducing false positives reduces overall healthcare costs.

描述（由申请人提供）：本申请涉及广泛的挑战领域生物标志物发现和验证（03）和特定挑战主题成像生物标志物（03-AR-104）。我们希望检测与皮肤癌发病相关的重要生物标志物，特别是黑色素瘤。具体来说，我们将应用一种新型成像技术（瞬态吸收显微镜）对固定皮肤和活体皮肤中发生的皮肤病变中的黑色素和血红蛋白进行成像。目标是无创地检测早期发展的黑色素瘤，并减少皮肤镜检查和组织病理学中的假阳性和假阴性。传统的黑色素瘤检测和诊断是一个两层过程，首先对可疑痣进行目视或皮肤镜检查，最后去除可疑组织，在显微镜下检查以确认诊断。然而，这种方法面临两个基本挑战。首先是难以通过肉眼辨别黑色素瘤和良性痣之间的差异；大多数黑色素瘤色素沉着，即使使用皮肤镜，医生也无法看到表面深处的情况。这个问题主要通过切除、H&E 染色和病理学家检查来解决，尽管患者通常会出现太多的痣，无法以这种侵入性方式切除和测试。第二个挑战是，即使对于训练有素的观察者来说，组织病理学的假阴性和假阳性仍然是一个严重的问题。假阴性会延误治疗并增加死亡率；事实上，黑色素瘤误诊是美国癌症医疗事故索赔的第二大常见原因（仅次于乳腺癌）。误报会通过不必要的、昂贵的和侵入性的程序（包括前哨淋巴结活检和全身辅助治疗）增加医疗成本，并可能使患者无法获得健康或人寿保险。因此，更准确的检测和诊断可能对患者生存和降低医疗成本产生非常大的影响。现有的显微镜方法不太适合应对这些挑战。反射共焦扫描激光显微镜 (rCSLM) 已在该应用中实现商业化，但对比度（散射）缺乏特异性，因为它不针对特定的生物标志物。传统的多光子显微镜可以以显微分辨率对组织深处进行成像，但色素病变是一个可怕的目标，因为产生的光被重新吸收（无论如何，来自黑色素的荧光非常弱）。在这里，我们使用非线性瞬态吸收显微镜，它不保留多光子显微镜的分辨率优势，但确实需要样品产生新波长的光。这种方法已经存在了几十年，但 PI 实验室最近的工作利用了先进的飞秒脉冲整形和脉冲序列调制方法来显着提高灵敏度，从而可以用适度的功率对组织进行成像。他和他的研究小组已将这项技术从基础科学转移到临床应用。靶向生物标志物对于黑色素瘤的诊断具有重要意义。有充分的证据表明，在发展中的黑色素瘤中，褐黑素和真黑素含量之间的局部比例发生了改变。在这里，我们通过在组织深处和非侵入性地显微镜下（在黑素体和黑素细胞中）测量这种分布，极大地改进了之前的工作。还有证据表明，微血管和氧合（氧合血红蛋白和脱氧血红蛋白，我们也可以对其进行无创深度成像）与转移潜力相关。总体而言，该项目的研究团队包括化学家、激光技术专家、病理学家和皮肤科医生，以便真正收集产生临床影响所需的专业知识。例如，PI是分子成像和超快激光物理学领域的先驱（也是美国物理学会激光科学分部的候任主席），同时也是化学、放射学和生物医学工程领域的教授。第一个目标是，我们专注于用我们的新方法和最好的传统方法分析去识别的切除痣。随着这项工作的发展，我们确定了与癌症发展最相关的分子特征，我们将进行新切除的皮肤研究，以验证安全功率限制（尽管我们目前使用的激光功率比现有的商业系统少）。最后，我们进展到活体动物模型（将诱导病变的人类皮肤移植到裸鼠上），我们还将测量血流量和局部组织氧合，作为侵袭性代谢的可能标志。到资助期结束时，我们将准备好对体内黑色素瘤的发展提供精彩而新颖的见解，并为人类临床工作做好准备。公共健康相关性：我们提出了一种新型成像技术，可以对正在发生的皮肤病变中的特定癌症生物标志物进行成像，无需切除即可无创地检测早期黑色素瘤，并减少组织病理学中的假阳性和假阴性。减少假阴性将减少癌症死亡率；减少误报可降低总体医疗成本。