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: 7640915
• 负责人: Oleg A Andreev
• 金额: $ 65.87万
• 依托单位: UNIVERSITY OF RHODE ISLAND
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-01 至 2013-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7640915
• 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）：我们的项目基于水溶性膜肽 pHLIP 的使用，我们通过全身荧光和 PET 成像表明，它可以选择性地靶向体内酸性实体瘤，并将极性货物分子转移到培养的癌细胞的细胞质中。 pHLIP 在弱酸性条件下作为单体单向插入细胞膜的脂质双层，如在肿瘤中发现的那样，并形成跨膜 α 螺旋，而在健康组织的正常细胞外 pH 值下，几乎没有插入细胞膜。迄今为止，尚未观察到 pHLIP 暴露对培养细胞或小鼠的毒性作用。在这里，我们建议开发一个基于 pHLIP-生物纳米注射器的纳米技术平台，用于选择性地将成像和治疗剂递送到肿瘤。通过将货物分子附着在 pHLIP 膜外的末端，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、治疗和治疗监测。通过可裂解的连接将货物连接到其插入端，pHLIP 可用于将细胞不可渗透的极性分子选择性易位到癌细胞中。通过三个实验室的共同努力，这项有前景的技术的广泛发展将成为可能。我们将使用 pHLIP 靶向来测试癌症模型，并确定肿瘤生长和发育与肿瘤酸度的关系。为了改进 pHLIP 技术，我们将设计、合成和测试各种树枝状-pHLIP 构建体，以便能够向肿瘤递送多种治疗和/或成像探针。我们将介绍一种通过可裂解的 S-S 键将货物分子和荧光染料同时缀合到 pHLIP C 末端的合成方案，并确定 pHLIP 可以通过膜的脂质双层易位的货物分子的特性（极性、形状、电荷和大小），从而定义可用于治疗肿瘤的新型极性治疗分子 治疗。我们将测试 pHLIP 在体内传递两种功能性细胞不可渗透分子的细胞内递送：毒素（鬼笔环肽）和基因调节剂（肽核酸）。重要的是，我们将尝试通过标记的 pHLIP-鬼笔环肽同时检测和治疗肿瘤，这是我们潜在抗转移药物的第一个先导。此外，我们将开发一种两步递送方案，用于体内癌细胞表面纳米颗粒的特异性束缚和组装：1）使用带有结合域的pHLIP靶向肿瘤，该结合域将被束缚到癌细胞表面；2）用含有治疗和/或成像有效负载并具有表面暴露的互补结合域的脂质体靶向pHLIP。受到当前版本 pHLIP 特性的启发，我们将进一步评估 pHLIP 序列变异对肽插入膜的影响，从而设计出具有一系列有用特性的第二代纳米注射器。 pHLIP 纳米技术为癌症的疾病特异性成像和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，癌症约占美国和其他发达国家所有死亡人数的 25%。我们的技术开发可以在这个问题的几个方面发挥作用，但主要概念是选择性地将治疗剂和成像剂递送到肿瘤细胞。该技术的另一个方面是，它允许使用一类新型治疗剂：细胞不可渗透的分子，只能转移到患病组织的细胞中，而不影响健康细胞。基于这些概念的疗法将表现出更高的功效和/或显着减少的副作用。这种改进对于癌症治疗尤其重要，因为大多数抗癌药物都是损害正常细胞的毒药。