Targeted Ceramic Nanovectors for Simultaneous Therapy and Imaging of Cancer

描述(申请人提供)：这项研究的总体目的是开发多功能核壳层状双氢氧化物纳米颗粒(LDHN)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为具有以下特征：1)金属或量子点核/LDH陶瓷外壳，用于成像所传递的纳米矢量和用于癌细胞的消融；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，以及3)用于靶向癌细胞的功能化最外表面。假设在一个生物相容的纳米级递送平台上整合靶向、成像和消融/化疗能力是前列腺癌治疗的一种有效方法，可直接应用于残留和转移疾病的治疗。无机陶瓷具有良好的生物相容性和在生理环境中的稳定性，是一种极具吸引力的生物应用材料。LDH纳米粒子是一类通式为M2 1-xM3 x(OH)2.(An-)x/n.mH2O的无机陶瓷，其中M2为二价阳离子，M3为三价阳离子，An-为n价层间阴离子。在目前的应用中，我们将使用基于核-壳LDH的纳米载体来靶向地将阴离子治疗药物(共轭阿霉素和抗bcl2 siRNA)输送到前列腺癌细胞。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。银核-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高温消融的附加属性。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有过度表达，因此适合于靶向恶性前列腺癌细胞。靶向PSMA的双分子(多肽和抗体)将与表面激活的LDHN偶联，以特异性靶向恶性前列腺细胞。该平台将在体外评估LDHN中的插层药物对前列腺癌细胞系和非恶性细胞的靶向性输送。我们的具体目标是：1)制备尺寸可控且尺寸分布窄的核-壳层状双氢氧化物纳米粒子(LDHN)；以半导体量子点和银纳米棒为核心同时进行成像和治疗；2.LDHN的表面活化，以及聚乙二醇(PEG)和前列腺靶向生物分子(PTB)在LDHN上的偶联；3)化疗分子在LDHN结构中的嵌入；3.体外评价以确定PTB-LDHN纳米载体的摄取、细胞内定位、有效性、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、分子和细胞生物学以及分子治疗和工程方面的互补经验，用于多功能核壳LDHN纳米载体的设计、生成、表征和体外评估，用于前列腺癌治疗。这项拟议研究的成功完成将为晚期前列腺癌疾病带来内置冗余和成像的新型靶向疗法，并可作为其他转移性癌症的治疗和成像的通用平台。 项目简介：我们合作研究的目标是开发有针对性的多功能核壳层状双氢氧化物(LDH)纳米矢量，用于晚期癌症的靶向破坏和光学成像。为此，我们开展了核壳结构LDH纳米粒子的设计、合成和表征工作。虽然核心将由荧光量子点或近红外(NIR)活性银(Ag)纳米棒组成，但层状双金属氢氧化物的外壳将有诱导细胞凋亡的基因毒素和嵌入其结构的siRNA。除了诱导细胞凋亡的分子外，在基于银纳米棒的核壳纳米粒子的情况下，加热消融也将促进癌细胞的破坏，从而导致双重治疗。肿瘤细胞靶向生物分子(多肽和抗体)将连接到LDH外壳的最外层，从而在单一输送平台上形成具有靶向、成像和双重治疗能力(消融和化疗)的多功能纳米载体。为了评价这些新型纳米载体的有效性，我们将利用人前列腺癌细胞系和未转化的前列腺上皮细胞，对核壳LDH纳米载体的细胞毒性、细胞死亡机制和选择性进行广泛的体外评估。

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.