Improved Peptide-based Tumor Targeting Agents Using Phage Display

使用噬菌体展示改进基于肽的肿瘤靶向剂

• 批准号: 7775087
• 负责人: SUSAN L DEUTSCHER
• 金额: $ 19.44万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-03-01 至 2012-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7775087
• 关键词: Adverse effects; Affinity; Amino Acids; Animals; Antibodies; Antigens; Avidity; Bacteriophages; Binding; Biodistribution; Biological Markers; Biological Models; Breast; Breast Carcinoma; Bypass; Cancer Detection; Cells; Charge; Consensus Sequence; Detection; Development; Dimerization; Diuretics; Dose; Epidermal Growth Factor Receptor; Foundations; Goals; Hepatobiliary; Human; Image; Imaging Techniques; Kidney; Label; Laboratories; Libraries; Life; Lung; Lung Neoplasms; Lysine; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of ovary; Mammary Neoplasms; Mission; Molecular Biology; Molecular Probes; Mus; Neoplasm Metastasis; Nude Mice; Ovarian; Ovary; Patient Care; Penetration; Peptide Phage Display Library; Peptides; Phage Display; Property; Proteins; Radiation; Radiolabeled; Radiopharmaceuticals; Research; Reticuloendothelial System; Signal Pathway; Staging; Study models; Surface; Techniques; Therapeutic Agents; Time; Toxic effect; Translations; Treatment Efficacy; Tumor Antigens; Urine; Work; base; cancer imaging; combinatorial chemistry; drug discovery; erbB-2 Receptor; improved; in vivo; innovation; kidney cell; neoplastic cell; novel; novel strategies; optical imaging; overexpression; particle; polypeptide; protein aminoacid sequence; public health relevance; radiotracer; research study; scaffold; single photon emission computed tomography; small molecule; tumor; tumor xenograft; tumorigenesis; uptake; vector

DESCRIPTION (provided by applicant): While treatable in their earliest stages, metastatic forms of breast, lung, and ovarian cancer are often lethal. Improved cancer survival will result from a more directed approach in which antigens overexpressed on tumors are targeted. Even though a limited number of tumor biomarkers are being exploited in antibody-based radiopharmaceutical drug discovery, there remains a critical need to identify new targeting vehicles and biomarkers. Peptide-based probes to facilitate cancer detection and therapy are rapidly evolving from combinatorial chemistry and bacteriophage (phage) display approaches. Many radiolabeled peptides and small proteins have shown good tumor-targeting propensity in vivo, however their translation into clinically useful agents has been slowed by their almost universal high renal uptake and retention. While the mechanisms for the observed uptake and retention are not clear, it is believed that positively charged peptides bind negatively charged kidney cells resulting in high doses of kidney radiation. Current techniques to block this retention, including lysine or diuretic administration are marginally effective and have harmful side effects. The overall goal of the proposed research is to employ innovative functional selection phage display approaches to improve the imaging and, potentially, therapeutic efficacy of tumor-targeting radiolabeled peptides. We hypothesize that in vivo phage display can be exploited to identify phage and corresponding peptides that are rapidly excreted in the urine and not retained in the kidneys or cleared through the reticuloendothelial system. These peptide sequences will be appended to tumor-targeting peptides and examined for their abilities to reduce renal retention. Phage display and affinity maturation studies will also be performed with a tumor-targeting micro-library, to generate peptides with improved tumor binding and reduced kidney retention in vivo. We have chosen as a model system the ErbB-2 receptor (ErbB-2)-targeting peptide, KCCYSL, discovered from phage display. Previous studies demonstrated that radiolabeled versions of KCCYSL bound the Erbb-2 receptor and imaged breast carcinomas in vivo, although high radiolabel kidney uptake and retention was observed. This well-characterized peptide will be the focus of these studies because it was originally identified in the context of phage and we know that the peptide retains its tumor targeting properties when it is expressed on phage or chemically synthesized. Furthermore, ErbB-2 overexpression and dimerization activates several signaling pathways that promote breast, lung, and ovarian tumorigenesis- making it an attractive target for development of new cancer imaging and therapeutic agents. The objectives of this proposed research are to: 1) develop novel strategies to select phage display libraries in non-tumor bearing mice in order to identify phage and corresponding peptides that are rapidly excreted into the urine; 2) in vivo affinity maturate 15 amino acid phage libraries displaying the ErbB-2 minimum binding motif, CCYSL, in breast tumor bearing mice to identify peptides with high tumor uptake and low kidney retention, and 3) examine the single photon emission computed tomography imaging efficacy of 111In-radiolabeled ErbB-2-targeting peptide constructs. This work is highly relevant to the mission of NCI and patient care in that the discovered peptides will form the foundation for new, well tolerated targeting agents for the detection and potential treatment of breast, ovary, and lung cancer. PUBLIC HEALTH RELEVANCE: The development of peptide and small molecule-based radiopharmaceutical for targeted cancer detection and therapy is hindered by the almost universal uptake and retention of peptides and small molecules in the kidneys. Our work is relevant in that new molecules will be discovered using highly innovative phage display techniques to reduce radiolabeled peptide uptake in the kidney by selecting phage and corresponding displayed peptides that bypass clearance through the reticuloendothelial system and are rapidly excreted in the urine. The ability of the discovered molecules to reduce kidney uptake of radiolabeled peptides that bind the ErbB-2 receptor overexpressed on breast, ovarian, and lung tumors will be investigated in living animals.

描述（由申请人提供）：虽然在早期阶段可以治疗，但乳腺癌、肺癌和卵巢癌的转移形式通常是致命的。提高癌症存活率将来自于更直接的方法，其中靶向肿瘤上过表达的抗原。尽管在基于抗体的放射性药物发现中利用了有限数量的肿瘤生物标志物，但仍然迫切需要鉴定新的靶向载体和生物标志物。促进癌症检测和治疗的基于肽的探针从组合化学和噬菌体（噬菌体）展示方法迅速发展。许多放射性标记的肽和小蛋白质在体内显示出良好的肿瘤靶向倾向，然而，它们转化为临床有用的药物的速度因其几乎普遍的高肾摄取和保留而减慢。虽然观察到的摄取和保留的机制尚不清楚，但认为带正电荷的肽结合带负电荷的肾细胞，导致高剂量的肾辐射。目前阻止这种滞留的技术，包括赖氨酸或利尿剂的施用，效果甚微，并且具有有害的副作用。这项研究的总体目标是采用创新的功能选择噬菌体展示方法来改善肿瘤靶向放射性标记肽的成像和潜在的治疗效果。我们假设体内噬菌体展示可以用来鉴定噬菌体和相应的肽，这些肽在尿液中迅速排泄，而不保留在肾脏中或通过网状内皮系统清除。这些肽序列将被附加到肿瘤靶向肽，并检查它们减少肾潴留的能力。还将用肿瘤靶向微文库进行噬菌体展示和亲和力成熟研究，以产生具有改善的肿瘤结合和减少的体内肾滞留的肽。我们选择了从噬菌体展示中发现的ErbB-2受体（ErbB-2）靶向肽KCCYSL作为模型系统。先前的研究表明，放射性标记的KCCYSL与Erbb-2受体结合，并在体内对乳腺癌进行成像，尽管观察到了高放射性标记的肾脏吸收和保留。这种充分表征的肽将成为这些研究的焦点，因为它最初是在噬菌体的背景下鉴定的，并且我们知道当它在噬菌体上表达或化学合成时，肽保留了其肿瘤靶向特性。此外，ErbB-2过表达和二聚化激活了促进乳腺、肺和卵巢肿瘤发生的几种信号通路，使其成为开发新的癌症成像和治疗药物的有吸引力的靶点。本研究的目的是：1）开发新的策略来选择非荷瘤小鼠中的噬菌体展示文库，以鉴定快速排泄到尿液中的噬菌体和相应的肽; 2）体内亲和成熟展示ErbB-2最小结合基序CCYSL的15个氨基酸噬菌体文库，在携带乳腺肿瘤的小鼠中鉴定具有高肿瘤摄取和低肾滞留的肽，和3）检查111 In-放射性标记的ErbB-2-靶向肽构建体的单光子发射计算机断层摄影成像功效。这项工作与NCI的使命和患者护理高度相关，因为所发现的肽将形成用于乳腺癌、卵巢癌和肺癌的检测和潜在治疗的新的、耐受性良好的靶向剂的基础。公共卫生关系：用于靶向癌症检测和治疗的基于肽和小分子的放射性药物的开发受到肽和小分子在肾脏中几乎普遍摄取和保留的阻碍。我们的工作是相关的，新的分子将被发现，使用高度创新的噬菌体展示技术，以减少放射性标记的肽在肾脏中的摄取，通过选择噬菌体和相应的展示肽，旁路清除通过网状内皮系统，并迅速排泄在尿液中。将在活体动物中研究所发现的分子降低放射性标记肽的肾脏摄取的能力，所述放射性标记肽结合在乳腺、卵巢和肺肿瘤上过表达的ErbB-2受体。

项目成果

Characterization of In Vivo Selected Bacteriophage for the Development of Novel Tumor-Targeting Agents with Specific Pharmacokinetics and Imaging Applications.

用于开发具有特定药代动力学和成像应用的新型肿瘤靶向剂的体内选择噬菌体的表征。

• DOI: 10.1007/978-1-4939-6911-1_29
• 发表时间: 2017
• 期刊: Methods in molecular biology (Clifton, N.J.)
• 作者: Newton-Northup,Jessica;Deutscher,SusanL
• 通讯作者: Deutscher,SusanL

In vivo bacteriophage peptide display to tailor pharmacokinetics of biological nanoparticles.

体内噬菌体肽展示可定制生物纳米颗粒的药代动力学。

• DOI: 10.1007/s11307-014-0762-z
• 发表时间: 2014
• 期刊: Molecular imaging and biology
• 影响因子: 3.1
• 作者: Newton-Northup,JessicaR;Dickerson,MarieT;Kumar,SenthilR;Smith,GeorgeP;Quinn,ThomasP;Deutscher,SusanL
• 通讯作者: Deutscher,SusanL