Targeted Ceramic Nanovectors for Simultaneous Therapy and Imaging of Cancer

描述（由申请人提供）：本研究的总体目的是开发基于核壳层状双氢氧化物纳米粒子（LDHN）的多功能纳米载体，用于晚期癌症疾病的靶向消融和成像。这种新颖的非聚合物平台将被设计成具有以下特征：1)金属或量子点核心/LDH陶瓷外壳，用于成像所传递的纳米载体和癌细胞的烧蚀；2)在陶瓷LDH外壳内嵌入诱导凋亡的化疗药物（基因毒素）或siRNA； 3)功能化的最外层表面用于靶向癌细胞。据推测，将靶向、成像和消融/化疗能力结合在一个单一的生物相容性纳米级递送平台上是前列腺癌治疗的一种强有力的方法，可直接应用于残余和转移性疾病的治疗。无机陶瓷由于其固有的生物相容性和在生理环境中的稳定性，在生物领域具有广泛的应用前景。LDH纳米颗粒是一类无机陶瓷，其通式为M2+1-xM3+x(OH)2.(An-)x/n。mH2O，其中M2+是二价阳离子，M3+是三价阳离子，An-是价n的层间阴离子。它们独特的结构很容易通过离子交换插入各种阴离子诱导凋亡的治疗药物。在目前的应用中，我们将采用核壳型ldh纳米载体，将阴离子疗法（结合阿霉素和抗bcl-2 siRNA）靶向递送到前列腺癌细胞。核心将分别由荧光半导体量子点和用于荧光和近红外（NIR）成像的银纳米棒组成。Ag核- ldh壳结构将具有通过使用纵向等离子体共振模式对癌细胞进行热消融的附加属性。前列腺特异性膜抗原（PSMA）是一种细胞表面受体，在前列腺癌疾病的所有阶段都过表达，因此适合靶向恶性前列腺癌细胞。靶向PSMA的双分子（多肽和抗体）将与表面活化的LDHNs结合，以特异性靶向恶性前列腺细胞。该平台将在体外进行评估，以靶向递送LDHN中的药物到前列腺癌细胞系和非恶性细胞。我们的具体目标是：1 .粒径可控、粒径分布窄的核壳层状双氢氧化物纳米粒子（LDHN）的合成与表征；以半导体量子点和银纳米棒为核心，用于同时成像和治疗；LDHN的表面活化，聚乙二醇（PEG）和前列腺靶向生物分子（PTBs）在LDHN上的偶联；3)化疗分子在LDHN结构内的嵌入；体外评价以确定PTB- LDHN纳米载体的摄取、细胞内定位、功效、选择性、消融机制和生物相容性。研究团队汇集了陶瓷材料工程，聚合物和表面化学，分子和细胞生物学，分子治疗和工程方面的互补经验，用于设计，生成，表征和体外评估多功能核-壳LDHN纳米载体作为前列腺癌治疗药物。该研究的成功完成将为晚期前列腺癌疾病带来具有内置冗余和成像的新型靶向治疗方法，并可作为其他转移性癌的治疗递送和成像的通用平台。

DESCRIPTION (provided by applicant): The overall purpose of this study is the development of multifunctional core-shell Layered Double Hydroxide Nanoparticle (LDHN)-based nanovectors for the targeted ablation and imaging of advanced cancer disease. This novel, non-polymeric platform will be designed to possess the following features: 1) metallic or quantum dot core/LDH ceramic shell for imaging the delivered nanovector and for ablation of cancer cells, 2) intercalated apoptosis-inducing chemotherapeutic drugs (genotoxins) or siRNA within the ceramic LDH shell, and 3) functionalized outermost surfaces for targeting cancer cells. It is hypothesized that the incorporation of targeting, imaging, and ablation/chemotherapeutic capabilities on a single biocompatible nanoscale delivery platform is a powerful approach for prostate cancer therapy, with direct application in the treatment of residual and metastatic disease. Inorganic ceramics are attractive materials for biological applications due to their inherent biocompatibility and stability in physiological environments. LDH nanoparticles are a class of inorganic ceramics that have a general formula of M2+1-xM3+x(OH)2.(An-)x/n.mH2O, where M2+ is a divalent cation, M3+ is a trivalent cation, and An- is the interlayer anion of valence n. Their unique structure readily allows the intercalation of a variety of anionic apoptosis-inducing therapeutics via ion exchange. In the present application, we will employ core-shell LDH-based nanovectors for the targeted delivery of anionic therapeutics (conjugated adriamycin and anti-bcl-2 siRNA) to prostate cancer cells. The core will consist of fluorescent semiconductor quantum dots and silver nanorods for fluorescent and near-infra red (NIR) imaging, respectively. The Ag core-LDH shell structure will have an additional attribute of hyperthermic ablation of cancer cells through the use of the longitudinal plasmon resonance mode. The Prostate-Specific Membrane Antigen (PSMA) is a cell-surface receptor over-expressed in all stages of prostate cancer disease and is therefore appropriate for targeting malignant prostate cancer cells. Bimolecular (peptides and antibodies) targeting the PSMA will be conjugated to surface-activated LDHNs in order to specifically target malignant prostate cells. The platform will be evaluated in vitro for the targeted delivery of the intercalated drugs in LDHN to prostate cancer cell lines and non-malignant cells. Our specific aims for this endeavor are: 1: Synthesis and characterization of core-shell layered double hydroxide nanoparticles (LDHN) with controlled size and narrow size distribution; with cores of semiconductor quantum dots and silver nanorods for simultaneous imaging and therapy, 2. Surface-activation of LDHN, and conjugation of poly(ethylene glycol) (PEG) and Prostate-Targeting Biomolecules (PTBs) on LDHNs, 3) Intercalation of chemotherapeutic molecules within the LDHN structure, and 3. In vitro evaluation to determine the uptake, intracellular localization, efficacy, selectivity, mechanisms of ablation, and biocompatibility of PTB- LDHN nanovectors. The team of investigators brings together complementary experience in ceramic materials engineering, polymer and surface chemistry, molecular and cellular biology, and molecular therapeutics and engineering, for the design, generation, characterization and in vitro evaluation of multifunctional core-shell LDHN nanovectors as prostate cancer therapeutics. Successful completion of the proposed research will result in novel targeted therapeutics with built-in redundancy and imaging for advanced prostate cancer disease, and can serve as a general platform for therapeutic delivery and imaging of other metastatic carcinomas. Project Narrative: The goal of our collaborative research is to develop targeted multifunctional core-shell Layered Double Hydroxide (LDH) nanovectors for the targeted destruction and optical imaging of advanced cancer disease. Towards that end, we carry out the design, synthesis, and characterization of core-shell LDH nanoparticles. While the core will consist of fluorescent quantum dots or near infra red (NIR) active silver (Ag) nanorods, the shell of layered double metal hydroxides will have apoptosis-inducing genotoxins and siRNA that are intercalated within its structure. In addition to apoptosis-inducing molecules, cancer cell destruction will also be facilitated by hyperthermic ablation in the case of Ag-nanorod based core-shell nanoparticles, resulting in dual therapy. Cancer cell targeting biomolecules (peptides and antibodies) will be conjugated to the outermost layer of the LDH shell leading to multifunctional nanovectors that possess targeting, imaging, and dual therapeutic capabilities (ablation and chemotherapy) on a single delivery platform. Extensive in vitro evaluation of cytotoxicity, mechanisms of cell death, and selectivity of the core-shell LDH nanovectors will be carried out using human prostate cancer cell lines and untransformed prostate epithelial cells in order to evaluate the efficacy of these novel nanovectors.