scVEGF Targeted Radiotherapy of Mammary and Colonic Cancer

scVEGF 乳腺癌和结肠癌靶向放疗

基本信息

批准号：
7847454
负责人：
FRANCIS Gerard BLANKENBERG
金额：
$ 33.28万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-05-24 至 2011-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7847454
关键词：
90Y Angiogenesis Inhibitors Autoradiography Biodistribution Biological Assay Blood Vessels Breast Adenocarcinoma Bystander Effect Capillary Endothelial Cell Cell Line Chemistry Clinical Trials Colon Carcinoma Colonic Neoplasms Controlled Study Cysteine Data Diagnostic Imaging Dose Electrons Endothelial Cells Engineering Ensure FDA approved Fibroblast Growth Factor Future Goals Human Hypoxia Image Imaging Techniques Isotopes Label Link Literature Location Maleimides Mammary gland Microscopic Modeling Mus N-terminal Neoplasm Metastasis Neoplasms in Vascular Tissue Palpable Patients Pharmaceutical Preparations Pharmacotherapy Primary Neoplasm Procedures Proteins Radiation Radiation therapy Radio Radiolabeled Recombinants Resistance Risk Schedule Site Surface Sutent Targeted Radiotherapy Techniques Testing Therapeutic Time Tissues Tracer Tumor Angiogenesis Tumor Volume VEGF121 gene Vascular Endothelial Cell Vascular Endothelial Growth Factor A Vascular Endothelial Growth Factor Receptor Vascular Endothelial Growth Factor Receptor-1 Vascular Endothelial Growth Factor Receptor-2 Vascular Endothelial Growth Factors Y 90 Ibritumomab Tiuxetan base cancer cell cell killing clinical application dimer dosimetry improved in vivo iodine-131-tositumomab killings malignant colon tumor neoplastic cell pre-clinical radiotracer receptor density research study response single photon emission computed tomography soft tissue success tumor tumor vascular supply uptake

项目摘要

The abundance of vascular endothelial growth factor (VEGF) receptors on surface of tumor vessel endothelial cells (ECs) is an attractive target for therapy. We propose to use a genetically engineered form of VEGF-A (VE0F121) labeled for radiotherapy of primary tumors in mice as well as for SPECT imaging for the serial non-invasive assessment of VEOF receptor density. This new protein combines the two 3-112 aa fragments of VEGFI2I (normally a dimer) fused into a single-chain (sc) with a N-terminal l5-aa tag containing a unique(C4) cysteine residue for site-specific labeling with simple, facile and non-destructive maleimide-based chemistry without byproducts or unmodified protein; problems that are inherent with other labeling techniques. scVEGF also has no pro-angiogenic domain thereby eliminating the risk of increasing tumor angiogenesis and the spread of metastases. Efforts to starve tumors of blood supply by targeting ECs have had mixed success and may be due to the unintended drug selection of resistant hypoxic tumor cell clones. We propose a new strategy which leverages the ability of scVEGF linked to isotopes to target tumor vessels and kill malignant cells directly. The most appropriate isotopes appear to be those with intense beta (Lu-177 or Y-90) or Auger electron emissions (In-Ill) that can kill cells within a small local range while sparing adjacent tissues. In this proposal we will model the size, geometry, location (with respect to tumor margins), and VEGF receptor density of the angiogenic rim over time in tumor bearing mice using biodistribution, autoradiographic of Lu-I77 (or Y-90) labeled scVEGF and immunohistochemical assays. These studies will be conducted with several well characterized cell lines; 411 syngeneic mammary, human MDA mammary and the human HT29 colonic carcinoma models. We will study both small (2-3 mm) and large tumors (6-9 mm) in all lines and with our lucifcrase expressing 41'I and MDA constructs we will also examine tumors that are non-palpable but can he seen with BLI imaging (very small < 1mm) to ascertain the relationships of the angiogenic rim to tumor size over time. From these data we will then perform dosimetry calculations and screen normal mice for signs ofradiotoxicity at the chosen tumor dose prior to actual radiotherapy. Short term, long-term (survival), and dose escalation radiotherapy studies will then be performed and compared with untreated (-) controls and Sutent (anti-angiogenic) treatment (+ controls). Successful completion of our proposal will give the needed preclinical data for pursuing scVEGF targeted radiotherapy in other models and ultimately clinical applications.

肿瘤血管内皮细胞（ECS）表面上的血管内皮生长因子（VEGF）受体的丰度是治疗的有吸引力的靶标。我们建议使用标记为小鼠原发性肿瘤的放射疗法的基因工程形式的VEGF-A（VE0F121），以及用于对VEOF受体密度的串行非侵入性评估的SPECT成像。这种新的蛋白质结合了vegfi2i（通常是二聚体）的两个3-112 AA片段，融合到单链（SC）中，带有N端L5-AA标签，其中包含独特的（C4）半胱氨酸残基，用于现场特异性标记，并具有简单的，易于促进的雄性乳腺癌的化学，而无需依赖性依赖性依赖性依赖性依赖性或不限化的蛋白质;其他标签技术固有的问题。 SCVEGF也没有促血管生成结构域，从而消除了增加肿瘤血管生成和转移扩散的风险。通过靶向EC饥饿的血液供应肿瘤的努力取得了成功，可能是由于耐药性低氧肿瘤细胞克隆的意外选择。我们提出了一种新策略，该策略利用了与同位素链接的SCVEGF靶向肿瘤血管并直接杀死恶性细胞的能力。最合适的同位素似乎是那些具有强烈β（LU-177或Y-90）或螺旋钻电子排放（IN-IN）的同位素，它们可以在较小的局部范围内杀死细胞，同时避免相邻的组织。在此提案中，我们将使用生物分布，LU-I77（或Y-90）标记的SCVEGF和Immunoyno组织化学化学化合物鉴定的肿瘤轴承小鼠的血管生成边缘的大小，几何，位置（相对于肿瘤边缘）和血管生成边缘的VEGF受体密度。这些研究将以几种良好的细胞系进行。 411乳腺乳腺，人类MDA乳腺和人类HT29结肠癌模型。我们将在所有线条中研究小（2-3毫米）和大肿瘤（6-9 mm），并且通过表达41'i和MDA构建体的荧光素酶，我们还将检查不容易PALPALPALP的肿瘤，但他是否可以通过BLI成像（非常小的<1mm）来确定与脉管均与肿瘤的关系的关系，以确定脉管均与肿瘤的关系。从这些数据中，我们将执行剂量测定计算，并在实际放疗之前在所选肿瘤剂量处进行剂量毒素的迹象。然后将进行短期，长期（生存）和剂量升级放射疗法研究，并将其与未经处理的（ - ）对照组和不良（抗血管生成）治疗（+对照组）进行比较。成功完成我们的提案将提供所需的临床前数据，以在其他模型和最终临床应用中追求SCVEGF的靶向放射疗法。