Extending the Luminescence Lifetime in Breast Cancer Diagnostics

生态旅游- 1159966 bossmannintellectual优点。发展在局部阶段识别乳腺癌的癌症诊断，结合手术中对肿瘤边界的准确识别，将是非常有利的，因为几乎所有乳腺癌的死亡率都发生在癌症转移之后。到2012年，美国国家癌症研究所预计将有约。23万新发乳腺癌病例和4万例死亡。乳腺癌的5年相对生存率从局部期的83%和局部期的98%下降到远处期的23%。许多蛋白酶在乳腺癌及其周围组织中过度表达。与免疫测定法相比，发光测定法具有三个数量级的敏感性。此外，发光测定法只能检测化学活性蛋白酶，而免疫测定法检测活性蛋白酶和酶原的混合物。在体外和体内蛋白酶检测中，阻碍发光法取得突破的主要障碍是血液，特别是组织的自身荧光。生物标本的自身荧光限制了光学癌症检测方法的灵敏度、特异性和统计再现性。这对于早期癌症（血液检查）和早期转移（体内）的检测尤其有效。使用目前最先进的癌症治疗方法，早期发现癌症仍然是降低癌症死亡率的唯一有效选择。自旋允许的能量转移在纳秒内进行，而自旋禁止的能量转移需要更多的时间（50到250纳秒）。血液和组织的自身荧光在不到25ns的时间内衰减。因此，在此之后，只有来自发光蛋白酶传感器的信号将是可见的。没有自身荧光将大大提高信噪比。它还将显著缩短体外和体内蛋白酶测量所需的时间。自旋禁止能量转移发生在钌-聚吡啶敏化剂和化学附着的菁染料之间。敏化剂-染料双极体将附着在铁/氧化铁核/壳纳米颗粒上。只要双极体附着在纳米粒子上，就会发生双极体发出的所有发光的强烈等离子体猝灭。diad和纳米颗粒之间的连接将由一致序列（寡肽）组成，该序列只能通过其各自的蛋白酶快速切割。多达四种不同的花青素染料，具有不同的吸收和发射最大值，将被用作双组件。当蛋白酶从纳米颗粒中释放出来时，每个二极管将表现出一个特征的、长寿命的发光信号（蛋白酶检测的光开关）。能够自旋禁止能量转移介导发光的纳米平台将在携带4T1肿瘤的BALB C小鼠中进行测试。4T1转移性乳腺癌模型是一种同基因模型。我们将对肿瘤位置进行成像，手术切除肿瘤，对切除的肿瘤组织进行表征，并进行肿瘤切除后的小鼠生存研究。核/壳铁/氧化铁NPs也可能用于MRI成像。更广泛的影响。意义和更广泛的影响：该项目的结果可能会导致一种新的、相对便宜的早期癌症诊断方法，以及乳腺癌手术的新护理标准。此外，该项目将提供有关癌症相关蛋白酶的活性如何根据体外和体内条件变化的定量数据。这些发现将对酶激活抗癌药物的开发具有重要意义。堪萨斯州立大学（KSU）解剖学、生理学和化学系将开设一门为期一周的单元课程，以提高学生的能力。生物光子学，生物传感和纳米医学的观点：Bossmann博士将开发和教授这门名为？开展?作为春季学期的一部分？材料化学课程？（chm852）。本课程面向兽医学、化学、生物化学、生物学和物理学等专业的研究生和本科生。拟议的研究将有助于将这项研究扩展到堪萨斯州立大学正在进行的癌症治疗和诊断方面的合作努力。这项研究也将为即将入学的本科生研究经验计划（REU）、本科生暑期研究机会计划（SUROP）和发展学者计划（DSP）创造机会。KSU的学生。拟议的工具开发将开放与其他大学和研究机构合作。

CBET-1159966BossmannIntellectual Merit. Developing cancer diagnostics for recognizing breast cancer at the localized stage, in combination with the exact identification of the tumor boundaries during surgery, would be very advantageous, because virtually all breast cancer mortality occurs after the cancer has metastasized. In 2012, the National Cancer Institute anticipates approx. 230,000 new breast cancer cases and 40,000 deaths. The 5-year relative survival of breast cancer decreases to 23 percent at the distant stage, from 83 percent at the regional stage and 98 percent at the localized stage. Numerous proteases are overexpressed in breast cancer and surrounding tissue. Luminescence assays have the potential to be three orders of magnitude more sensitive, when compared to immunoassays. Furthermore, luminescence assays are able to detect only chemically active proteases, whereas immunoassays detect a mixture of active proteases and zymogens. The major impediment that has prevented the breakthrough of luminescence assays for in-vitro and in-vivo protease detection is the autofluorescence of blood and especially tissue. Autofluorescence of biospecimens limits the sensitivity, specificity and statistical reproducibility of optical cancer detection methods. This is especially valid for the detection of early cancers (in blood tests) and early metastases (in-vivo). Using the current state-of-the-art cancer treatment, early detection of cancer remains the only valid option for decreasing cancer mortality. Whereas spin-allowed energy transfer proceeds within nanoseconds, spin-forbidden energy transfer requires significantly more time (50 to 250 ns). The autofluorescence of blood and tissue decays within less than 25ns. Therefore, only the signal from the luminescent protease sensor will be visible after that time. The absence of autofluorescence will greatly enhance the signal-to-noise ratio. It will also significantly shorten the time required for in-vitro and in-vivo protease measurements. Spin-forbidden energy transfer occurs between a ruthenium-poly-pyridyl sensitizer and a chemically attached cyanine-dye upon excitation of the sensitizer. The sensitizer-dye diad will be attached to iron/iron oxide core/shell nanoparticles. Intense plasmon quenching of all luminescence occurring from the diad will occur as long as the diad is attached to the nanoparticle. The linker between diad and nanoparticle will consist of a consensus sequence (oligopeptide) that can be cleaved quickly only by its respective protease. Up to four different cyanine dyes featuring different absorption and emission maxima will be used as components in a diad. Upon release by a protease from the nanoparticle, each diad will exhibit a characteristic, long-lived luminescence signal (Light Switch for Protease Detection). The nanoplatforms capable of spin-forbidden energy transfer mediated luminescence will be tested in BALB C mice bearing 4T1 tumors. The 4T1 metastatic breast cancer model is a syngeneic model. We will image the tumor location, surgically excise the tumors, characterize the excised tumor tissue and conduct a mouse survival study after tumor excision. The core/shell iron/iron oxide NPs can also potentially be used for MRI imaging.Broader Impacts. Significance and Broader Impact: The results from the project could potentially lead to a new and relatively inexpensive method for early cancer diagnostics and a new standard of care in breast cancer surgery. In addition, the project will provide quantitative data with respect to how the activity of cancer-related proteases varies depending on in-vitro and in-vivo conditions. These findings will be of importance for the development of enzyme-activatable anticancer drugs. The Departments of Anatomy&Physiology and Chemistry at Kansas State University (KSU) will create one block course for the duration of one week in order to enhance their students? perspective on biophotonics, biosensing and nanomedicine: Dr. Bossmann will develop and teach this block course entitled ?Theranostics? during the spring semester as a part of the ?Materials Chemistry Course? (CHM 852) of Chemistry at KSU. This course is open to all graduate and undergraduate students in veterinary medicine, chemistry, biochemistry, biology and physics. The proposed study will help extend this research to ongoing collaborative efforts on cancer therapy and diagnostics at Kansas State University. The research will also create opportunities to incoming REU (Research Experience of Undergraduates), SUROP (Summer Undergraduate Research Opportunity Program) and DSP (Developing Scholars? Program) students at KSU. The proposed instrumental development will be open to collaborations with other Universities and Research Institutions.