pHLIP Nanotechnology Platform for Cancer Imaging and Therapy

用于癌症成像和治疗的 pHLIP 纳米技术平台

• 批准号: 7910974
• 负责人: Oleg A Andreev
• 金额: $ 58.16万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7910974
• 关键词: Acidity; Acidosis; Adenocarcinoma; Adverse effects; Affect; Amino Acids; Animals; Antineoplastic Agents; Binding; Biodistribution; Cancer Model; Cancerous; Cell membrane; Cell surface; Cells; Cessation of life; Charge; Cleaved cell; Cyclic Peptides; Cytoplasm; Detection; Developed Countries; Developing Countries; Development; Diagnosis; Diagnostic Neoplasm Staging; Disease; Dyes; Endocytosis; Exhibits; Fluorescence; Fluorescent Dyes; Gene Expression Regulation; Generations; Goals; Growth and Development function; Hydrophobicity; Image; Implant; Inflammation; Kidney Neoplasms; Label; Laboratories; Lead; Link; Lipid Bilayers; Liposomes; Liver; Luciferases; Mediating; Membrane; Metabolism; Methods; Modeling; Monitor; Mus; Nanotechnology; Needles; Normal Cell; Organ; Pathway interactions; Peptide Nucleic Acids; Peptides; Pharmaceutical Preparations; Poison; Positron-Emission Tomography; Property; Receptor Cell; Scheme; Shapes; Signal Transduction; Site; Solid; Solid Neoplasm; Staging; Surface; Syringes; Technology; Testing; Therapeutic; Therapeutic Agents; Time; Tissues; Toxic effect; Toxin; Tumor stage; Variant; Vesicle; Water; alpha helix; base; cancer cell; cancer imaging; cancer therapy; design; extracellular; imaging probe; improved; in vivo; monomer; nanoparticle; novel strategies; protonation; technology development; tumor; tumor growth; water solubility

DESCRIPTION (provided by applicant): Our project is based on the use of a water-soluble membrane peptide, pHLIP, which we have shown, by whole-body fluorescence and PET imaging, to selectively target acidic solid tumors in vivo and to translocate polar cargo molecules into the cytoplasms of cultured cancer cells. pHLIP inserts unidirectionally across the lipid bilayer of a cell membrane as a monomer under mildly acidic conditions, as are found in tumors and forms a transmembrane alpha helix, whereas there is practically no insertion across the membranes of cells with the normal extracellular pH of healthy tissue. To date, no toxic effects of pHLIP exposure have been observed either for cells in culture or for mice. Here we propose to develop a nanotechnology platform for selective delivery of imaging and therapeutic agents to tumors based on the use of the pHLIP-bionanosyringe. By attaching cargo molecules to the end of pHLIP that stays outside of the membrane, we can anchor imaging or therapeutic probes to the surfaces of cancer cells, facilitating diagnosis, treatment and therapeutic monitoring. By attaching cargo to its inserted end via cleavable links, pHLIP can be used for the selective translocation of polar, cell-impermeable molecules into cancer cells. By combining the efforts of three laboratories, a broad development of this promising technology will be possible. We will use pHLIP targeting to test cancer models and establish how tumor growth and development correlate with tumor acidity. To improve pHLIP technology, we will design, synthesize and test various dendrimeric-pHLIP constructs to enable delivery of multiple therapeutic and/or imaging probes to tumors. We will introduce a synthetic scheme of simultaneous conjugation of cargo molecules and fluorescent dyes to the C-terminus of pHLIP via a cleavable S-S bond and establish the properties (polarity, shape, charge and size) of cargo molecules that pHLIP can translocate through the lipid bilayer of a membrane, defining a new, polar class of therapeutic molecules that can be delivered for tumor treatment. We will test pHLIP for the intracellular delivery of two functional cell-impermeable molecules in vivo: a toxin (phalloidin) and a gene regulation agent (Peptide Nucleic Acid). Importantly, we will attempt the simultaneous detection and treatment of tumors by labeled pHLIP-phalloidin, which is our first lead for a potential antimetastatic drug. Further, we will develop a two-step delivery scheme for the specific tethering and assembly of nanoparticles at the surfaces of cancer cells in vivo: 1) targeting tumors using pHLIP with a binding domain, which will be tethered to the surface of cancer cells and 2) targeting the pHLIP with liposomes containing therapeutic and/or imaging payloads and having a surface-exposed complementary binding domain. Inspired by the properties of pHLIP in its current version, we will further evaluate the effect of pHLIP sequence variation on peptide insertion into a membrane, enabling the design of a second generation of the nanosyringe with a range of useful properties. pHLIP nanotechnology offers a new approach for the disease-specific imaging and treatment of cancers. Our ultimate goal is to improve the diagnosis and treatment of cancer, which is responsible for about 25% of all deaths in the USA and other developed countries. There are several aspects of the problem where our technology development could be useful, but the major concept is the selective delivery of therapeutic and imaging agents to cells in tumors. Another aspect of the technology is that it permits the use of a new class of therapeutic agents: cell-impermeable molecules that would be translocated into cells only in diseased tissue while not affecting healthy cells. A therapy based on these concepts would exhibit much higher efficacy and/or significantly reduced side effects. Such improvements are especially important for cancer treatment, since the majority of anti-cancer drugs are poisons that damage normal cells.

描述（由申请人提供）：我们的项目基于使用水溶性膜肽Phlip的使用，我们通过全身荧光和PET成像显示，它在体内有选择地靶向酸性实体瘤并转化极性货物分子进入培养的癌细胞的细胞肿瘤。 phlip在细胞膜的脂质双层上单向插入在轻度酸性条件下作为单体，如在肿瘤和形成跨膜alpha螺旋中的单体，而实际上在细胞膜上没有插入正常的细胞膜的膜膜，没有插入。 。迄今为止，对于培养物或小鼠的细胞，尚未观察到phlip暴露的毒性作用。在这里，我们建议开发一个纳米技术平台，以根据使用phlip-bionanosyringe的使用选择性地递送成像和治疗剂向肿瘤。通过将货物分子连接到留在膜外的PHLIP末端，我们可以将成像或治疗探针锚定在癌细胞表面，从而促进诊断，治疗和治疗性监测。通过将货物连接到其插入的末端，可以将Phlip用于选择性易位，可渗透细胞的分子到癌细胞中。通过结合三个实验室的努力，这项有前途的技术将有可能。我们将使用PHLIP靶向测试癌症模型，并确定肿瘤生长和发育与肿瘤酸度的相关性。为了提高PHLIP技术，我们将设计，合成和测试各种树突型镀键型构建体，以使多种治疗和/或成像探针能够递送到肿瘤中。我们将通过可裂解的S-S-S键同时引入货物分子和荧光染料的综合方案，并建立Phlip可以通过脂质转运的货物分子的特性（极性，形状，形状，电荷和大小）膜的双层，定义了一种可以用于肿瘤治疗的新的极性治疗分子。我们将测试PHLIP在体内的细胞内递送两个功能性细胞的分子：A毒素（腓骨）和基因调节剂（肽核酸）。重要的是，我们将尝试通过标记的phlip-phalloidin同时检测和治疗肿瘤，这是我们的第一个潜在抗转移性药物的铅。此外，我们将开发一个两步递送方案，用于体内纳米颗粒在癌细胞表面的特定束缚和组装：1）使用PHLIP与结合结构域靶向肿瘤，将其链接到癌细胞和癌细胞表面和2）用含有治疗和/或成像有效载荷的脂质体靶向pHLIP，并具有表面暴露的互补结合结构域。受PHLIP在当前版本中的特性的启发，我们将进一步评估phlip序列变化对肽插入膜中的影响，从而使第二代纳米风格的设计具有一系列有用的特性。 Phlip纳米技术为疾病特异性成像和癌症治疗提供了一种新方法。我们的最终目标是改善癌症的诊断和治疗，这造成了美国和其他发达国家所有死亡人数的25％。问题的几个方面可以有用，但是主要概念是将治疗和成像剂的选择性输送到肿瘤中的细胞。该技术的另一个方面是，它允许使用一种新的治疗剂：可耐细胞的分子，这些分子仅在患病的组织中仅在不影响健康细胞的情况下转移到细胞中。基于这些概念的治疗将表现出更高的功效和/或显着降低副作用。这种改善对于癌症治疗尤为重要，因为大多数抗癌药物是损害正常细胞的毒药。