Extending the Luminescence Lifetime in Breast Cancer Diagnostics

延长乳腺癌诊断中的发光寿命

• 批准号: 1159966
• 负责人: Stefan Bossmann
• 金额: $ 30.46万
• 依托单位: Kansas State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-08-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1159966&HistoricalAwards=false
• 关键词: Extending; Luminescence; Lifetime; Breast; Cancer

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.

CBET-1159966博士曼智能优点。发展癌症诊断学，以便在局部阶段识别乳腺癌，并结合手术中准确识别肿瘤边界，将是非常有利的，因为几乎所有乳腺癌死亡都发生在癌症转移之后。2012年，美国国家癌症研究所预计大约会。新增乳腺癌病例23万例，死亡4万例。乳腺癌的5年相对生存率从区域阶段的83%和局部阶段的98%下降到远程阶段的23%。大量的蛋白水解酶在乳腺癌及其周围组织中过度表达。与免疫分析相比，发光分析的灵敏度有可能提高三个数量级。此外，发光检测只能检测化学活性的蛋白水解酶，而免疫检测检测活性蛋白和酵素的混合物。阻碍发光法在体外和体内检测蛋白酶方面取得突破的主要障碍是血液特别是组织的自身荧光。生物样品的自体荧光限制了光学癌症检测方法的敏感性、特异性和统计重复性。这对于检测早期癌症(血液检测)和早期转移(体内)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明显更多的时间(50到250 ns)。血液和组织的自发荧光在不到25 ns的时间内衰减。因此，在该时间之后，只有来自发光蛋白酶传感器的信号将是可见的。没有自发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。在Ru-聚吡啶敏化剂和化学结合的菁染料之间，在敏化剂的激发下发生自旋禁止的能量转移。敏化剂-染料二聚体将附着在铁/氧化铁核/壳纳米粒子上。只要二聚体附着在纳米颗粒上，就会发生强烈的等离子体猝灭来自二聚体的所有发光。二聚体和纳米颗粒之间的连接物将由一个共同的序列(寡肽)组成，只有它们各自的蛋白酶才能快速切割该序列。最多四种具有不同吸收和发射最大值的不同菁染料将被用作DIAD的成分。当酶从纳米颗粒中释放出来时，每个二聚体都将显示出一个特征的、长寿命的发光信号(用于检测蛋白水解酶的光开关)。能够自旋禁止能量转移介导的发光的纳米平台将在携带4T1肿瘤的BALB C小鼠身上进行测试。4T1转移性乳腺癌模型是一种同基因模型。我们将成像肿瘤的位置，手术切除肿瘤，确定切除的肿瘤组织的特征，并在肿瘤切除后进行小鼠存活研究。核壳结构的铁/氧化铁纳米粒子也有可能用于核磁共振成像。意义和更广泛的影响：该项目的结果可能会带来一种新的、相对廉价的癌症早期诊断方法，以及乳腺癌手术的新护理标准。此外，该项目将提供与癌症相关的蛋白酶活性如何随体外和体内条件而变化的定量数据。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街区课程，以提高他们的学生？生物光子学、生物传感和纳米医学的前景：博斯曼博士将开发和教授这门名为？Theranostics？春季学期作为材料化学课程的一部分。科罗拉多州立大学化学(CHM 852)本课程面向所有兽医、化学、生物化学、生物和物理专业的研究生和本科生。这项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。这项研究还将为REU(本科生的研究经验)、SUROP(暑期本科生研究机会计划)和DSP(发展中的学者？)创造机会。计划)学生在KSU。拟议的工具性开发将向与其他大学和研究机构合作开放。