Labeling nanobodies with 18F residualizing labels for HER2 specific PET imaging

使用 18F 残留标签标记纳米抗体，用于 HER2 特异性 PET 成像

基本信息

批准号：
8891591
负责人：
GANESAN VAIDYANATHAN
金额：
$ 36.3万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-01 至 2020-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8891591
关键词：
Accounting Adhesions Adverse effects Affinity Animal Model Antibodies Autopsy Binding Biodistribution Biological Assay Breast Cancer Cell Breast Cancer Patient Breast Cancer cell line Breast Carcinoma Catabolism Cell Line Cells Characteristics Chemistry Detection Development Diagnostic Neoplasm Staging Drug Targeting ERBB2 gene Early Diagnosis Economics Effectiveness Epidermal Growth Factor Receptor Exclusion Fab Immunoglobulins Family member Fluorescent in Situ Hybridization Fluorine Glycoproteins Goals Guanidines Half-Life High Pressure Liquid Chromatography Image Imaging Device Immunoglobulin G Immunohistochemistry In Vitro Kinetics Label MCF7 cell Malignant Neoplasms Medical Oncology Methods Micrometastasis Modeling Molecular Target Molecular Weight Monitor Mus Myocardial dysfunction Normal tissue morphology Oncogene Proteins Patients Peptides Positron-Emission Tomography Process Property Prosthesis Proteins Radioactive Iodine Radiochemistry Radioisotopes Radiolabeled Reagent Recurrence Regimen Relative (related person)Research Risk Role SKBR3 Signal Pathway Therapeutic Time Trastuzumab Woman Xenograft procedure accurate diagnosis analog base cancer cell cancer stem cell cell growth cost design gel electrophoresis imaging agent imaging modality immunoreactivity in vivo malignant breast neoplasm microPET migration molecular imaging nanobodies neoplastic cell novel overexpression personalized medicine phase I trial protein degradation public health relevance radiochemical radiotracer receptor research study response subcutaneous targeted treatment tumor uptake vector

项目摘要

DESCRIPTION (provided by applicant): HER2 is overexpressed in 25-30% of breast cancers and is a major contributor to tumor aggressiveness and recurrence. This has led to the development of HER2-targeted therapies such as trastuzumab, which can be effective in patients overexpressing HER2. Accurate assessment of tumor HER2 levels in real time could help personalize HER2-targeted therapy and monitor therapy response. Unlike current methods for assessing HER2 levels such as immunohistochemistry and fluorescence in situ hybridization (FISH), quantitative positron emission tomography (PET) imaging is noninvasive, can be performed repeatedly, and could provide accurate levels of HER2 in real time. Moreover, the high sensitivity of PET imaging could permit the detection of occult micrometastases and possibly HER2-expressing breast cancer stem cells. Nanobodies (Nbs) are single domain antibodies with a tenth of the molecular weight of an IgG, making them an ideal targeting vector for use in combination with the most widely utilized PET radionuclide, 18F. Conventional methods for protein labeling may result in the loss of 18F from tumor cells after a protein undergoes internalization and intracellular catabolism. We plan to extend the residualizing label (RL) approach, used successfully with radioiodine, to synthesize novel reagents designed to generate 18F-labeled catabolites that will be trapped in tumor cells after Nb internalization. Fluorine-18- labeled analogues of N-succinimidyl 4-guanidinomethyl-3-iodobenzoate (SGMIB), the best residualizing agent that we developed for radioiodination of internalizing biomolecules, will be generated. We seek to combine a new HER2 targeted 2Rs15d Nb developed by our colleague in Brussels, with a novel 18F- labeling chemistry with the goal of generating a PET imaging tool for assessing HER2 levels in breast cancer and thereby for the personalization of HER2 targeted therapy. If successful, the 18F-labeling reagents and methods that we shall develop could be applied to other proteins and peptides targeting HER2 and other internalizing receptors that are over expressed on breast cancers. We propose the following specific aims: 1. Develop prosthetic groups for 18F-labeling of Nbs based on the best Nb radioiodination residualizing agent, N-succinimidyl 4-guanidinomethyl-3-[*I] iodobenzoate ([*I]SGMIB). 2. Label Nbs with these agents and determine the characteristics of the labeled Nb conjugates. 3. Evaluate labeled Nbs in vitro using HER2-expressing breast cancer cells. 4. Evaluate labeled Nbs in vivo in mice bearing breast carcinoma xenografts.

描述（由申请人提供）：HER2 在 25-30% 的乳腺癌中过度表达，是肿瘤侵袭性和复发的主要因素，这导致了曲妥珠单抗等针对患者的 HER2 靶向疗法的开发。与目前评估 HER2 水平的方法不同，实时准确评估肿瘤 HER2 水平有助于个性化 HER2 靶向治疗并监测治疗反应。与免疫组织化学和荧光原位杂交（FISH）一样，定量正电子发射断层扫描（PET）成像是无创的，可以重复执行，并且可以实时提供准确的HER2水平，而且PET成像的高灵敏度可以进行检测。隐匿性微转移和可能表达 HER2 的乳腺癌干细胞的纳米抗体 (Nbs) 是单域抗体，其分子量仅为 IgG 的十分之一，使其成为理想的靶向载体。与最广泛使用的 PET 放射性核素 18F 结合使用，蛋白质标记的传统方法可能会在蛋白质经历内化和细胞内分解代谢后导致肿瘤细胞丢失 18F。成功地与放射性碘一起使用，合成了旨在产生 18F 标记的分解代谢物的新型试剂，这些代谢物在 Nb 内化后将被捕获在肿瘤细胞中。 N-琥珀酰亚胺基 4-胍基甲基-3-碘苯甲酸酯 (SGMIB) 的类似物是我们开发的用于内化生物分子放射性碘化的最佳残留剂，我们寻求结合我们在布鲁塞尔的同事开发的新的 HER2 靶向 2Rs15d Nb，使用新型 18F 标记化学，目标是生成 PET 成像工具，用于评估乳腺癌中的 HER2 水平，从而如果成功，我们将开发的 18F 标记试剂和方法可应用于针对乳腺癌中过度表达的 HER2 和其他内化受体的其他蛋白质和肽。 1. 基于最好的Nb放射性碘标记残留剂N-琥珀酰亚胺基，开发用于18F标记Nbs的辅基4-胍基甲基-3-[*I] 碘苯甲酸酯 ([*I]SGMIB) 2. 用这些试剂标记 Nb，并确定标记的 Nb 缀合物的特性 3. 使用表达 HER2 的乳腺癌细胞在体外评估标记的 Nb。 4. 在携带乳腺癌异种移植物的小鼠体内评估标记的 Nb。