Luminscent Dendrimer-Lanthanide Nanotechnology for Multiplex Detection

描述(申请人提供)：总体目标是将用于多重检测的基于镧系元素检测的纳米技术与肿瘤靶向多肽相结合，以创新癌症检测领域，并促进对实体肿瘤的靶向输送。这项技术的发展需要将对肿瘤生物学的理解与纳米技术化学相结合，并与NCI重点关注的纳米技术倡议保持一致。NCI的这种强调承认了通过合作方法和技术融合可以对癌症诊断和治疗产生的积极影响。这项提议将导致1)新型发光树枝状大分子-稀土纳米技术的开发，其原理是树枝状大分子支架的纳米尺寸可以解决并改进目前发光稀土报告的局限性，以及2)这种纳米级分子在癌症小鼠模型中用于多重检测的概念证明。多重检测将通过合成带有肿瘤血管靶向多肽外环的稀土配位的PAMAM(聚酰胺胺)树枝状大分子支架来实现，从而得到携带稀土阳离子的多肽-树枝状大分子(Ln-Pd)。该方案将利用Ln-Pd的独特发光特性(长寿命、独特的特征和尖锐的发射波段)来实现对自然生物自发荧光背景的灵敏和区分检测。通过创新地使用连接在PAMAM支链末端的敏化剂，可以获得强的稀土纳米分子发射信号；这是一种通过增加对激发光能的吸收并将其转移到几个内部定位的稀土元素来克服低量子产率的方法。反过来，镧系元素会发出强烈且独特的光谱信号。在体外表征和体外研究验证后，Ln-PD(通过RGD和RRL多肽靶向肿瘤血管)将在SCC SF/VII小鼠鳞状细胞癌模型中进行多重应用。该提案的成功完成将展示基于镧系元素的检测系统的功能性质，这可以通过改进发光显微镜和生物测试中的应用来实现。因此，检测肿瘤及其微环境的灵敏度的任何提高，都将对科学发现能力和临床医学的改进产生积极的影响。 该项目旨在利用树枝状大分子的纳米技术，通过创新地使用化学修饰树枝状大分子来改进对癌症的检测和传递。此外，树枝状大分子上还会附着一些小蛋白，以检测树枝状大分子与肿瘤模型系统结合的能力。

DESCRIPTION (provided by applicant): The overarching goal is to merge lanthanide detection-based nanotechnology for multiplex detection with tumor-targeting peptides to innovate in the field of cancer detection and to facilitate targeted delivery to solid tumors. The development of this technology requires the merging of an understanding of tumor biology with nanotechnology chemistry, and is aligned with the focused nanotechnology initiative of the NCI. Such emphasis by the NCI acknowledges the positive impact on cancer diagnosis and treatment that can be gained through collaborative approaches and the merging of technologies. This proposal will result in 1) the development of novel luminescent dendrimer-lanthanide nanotechnology on the principle that the nanoscale size of a dendrimer scaffold can address and improve upon the limitations of current luminescent lanthanide reporters and 2) a proof of concept demonstration of the utility of such nanoscale molecules for multiplex detection in a mouse model of cancer. The multiplex detection will be realized through synthesizing lanthanide-coordinating PAMAM (polyamidoamine) dendrimer scaffolds with an outer ring of tumor vasculature-targeting peptides; resulting in peptide-dendrimer molecules carrying lanthanide cations (Ln-PD). This proposal will exploit the unique luminescent properties of Ln-PD (long life, unique signatures, and sharp emission bands) to allow sensitive and discriminating detection against the background of natural biologic autofluorescence. The strong emission signal of lanthanide-nanomolecules is possible through the innovative use of sensitizers attached to the branch termini of the PAMAM; an approach which overcomes low quantum yield by increasing the absorption of excitation light energy and transferring it to several internally located lanthanides. In turn, the lanthanides emit a strong and spectroscopically unique signal. After characterizing in vitro and verifying with ex vivo studies, the Ln-PD (targeted to the tumor vasculature by peptides RGD and RRL) will be quantified in a multiplex application in the SCC SF/VII mouse model of squamous cell cancer. Successful completion of the proposal will demonstrate the functional nature of lanthanide-based detection systems which may by realized through improved applications in luminescence microscopy and bioassays. Subsequently, any improvement in the sensitivity of detection of tumors and their microenvironments will positively impact the ability of scientific discovery and improvements in clinical medicine. PUBLIC HEALTH RELEVANCE This project aims to use nanotechnology of dendrimers to improve the detection of and delivery to cancers through the innovative use of chemistry to modify the dendrimers. Additionally, small proteins will be attached to the dendrimers to allow assay of ability of the dendrimers to home to tumor model systems.