Targeting Pancreatic Cancer with Aptamers to the CCK-B Receptor

使用 CCK-B 受体适体靶向胰腺癌

• 批准号: 8644259
• 负责人: Gail L. Matters
• 金额: $ 16.14万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-02 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8644259
• 关键词: Affinity; Amino Acids; Animals; Antibodies; Apoptosis; Binding; Binding Sites; Biological Assay; Biological Availability; Biology; Bromodeoxyuridine; Cancer Research Project; Cell physiology; Cholecystokinin; Cholecystokinin B Receptor; Commit; DNA; Development; Diagnosis; Disease; Dose; ERBB2 gene; Epidermal Growth Factor Receptor; Evolution; Flow Cytometry; G-Protein-Coupled Receptors; Gastrins; Genetic; Goals; Growth; Human; In Vitro; Inhibitory Concentration 50; Injury; Intraepithelial Neoplasia; Label; Lesion; Libraries; Ligands; Link; Malignant Neoplasms; Malignant neoplasm of gastrointestinal tract; Malignant neoplasm of pancreas; Mediating; Mediation; Molecular; Monoclonal Antibodies; Mus; Mutation; Normal tissue morphology; Oligonucleotides; Pancreas; Pathologist; Patients; Peptides; Pharmaceutical Preparations; Play; Protein Tyrosine Kinase; Radioactive; Radiolabeled; Research; Role; Scientist; Signal Pathway; Site; Solid Neoplasm; Specificity; Structure; Survival Rate; Techniques; Testing; Therapeutic; Time; Toxic effect; aptamer; autocrine; cancer therapy; design; extracellular; gastrointestinal; immunogenic; improved; in vivo; innovation; oncology; outcome forecast; pancreatic cancer cells; pancreatic neoplasm; peripheral blood; public health relevance; radiotracer; receptor; receptor binding; screening; tumor specificity; uptake

DESCRIPTION (provided by applicant): Advances in cancer therapy have all come from understanding the biology and or genetic make-up of a particular cancer. When these factors are identified, treatments can then be 'personalized' with tumor specificity by targeting of cancer-related receptors, and less toxicity from off-site injury. Unlike other solid tumors, G-protein coupled receptors (GPCRs) have been shown to play a critical role in mediation of growth of gastrointestinal cancers. We have identified a receptor on pancreatic cancer, the CCK-B receptor, which is over-expressed in cancer compared to normal tissues and responsible for growth stimulation. Our research team has shown that gastrin stimulates growth of pancreatic cancer by mediating its effects in an autocrine fashion through this GPCR. We have shown that if gastrin's interaction at the receptor can be blocked, pancreatic cancer is inhibited. One problem has been in developing an agent to selectively block this receptor that can potentially be used therapeutically. Our lab and others have attempted to raise a monoclonal antibody to the gastrin binding site of the CCK-B receptor but have been unsuccessful due to the similar homology between human and mouse sequences. Therefore, we have undertaken a different strategy using DNA aptamers to target the CCK- B receptor in pancreatic cancer. DNA aptamers - highly structured oligonucleotides that bind selectively to receptor targets with affinities comparable to or better than antibodies, are more stable in the peripheral blood than monoclonal antibodies, and they are not immunogenic. We synthesized two peptides corresponding to the gastrin binding site on the extracellular, N-terminus of the CCK-B receptor. Using a technique called Systematic Evolution of Ligands by Exponential Enrichment (SELEX) we identified specific DNA aptamers that bind to the external gastrin binding site of the CCK-B receptor. We hypothesize that selective DNA aptamers to the CCK-B receptor can specifically target pancreatic cancer and inhibit growth by blocking the actions of gastrin. In order to test this hypothesis we propose the following aims: 1) Characterize the binding affinities (Kd) and IC50 of a panel of aptamers we have identified using pancreatic cancer cells in vitro to select aptamers with the greatest affinities for growth studies. 2) Study the efficacy of aptamers to inhibit growth of pancreatic cancer in culture and in mice bearing an orthotopic pancreatic cancer. The role of CCK-B targeted aptamers on apoptosis will also be investigated. The long term goal of our research is to improve treatment and survival of patients with pancreatic cancer.

描述(由申请人提供)：癌症治疗的进步都来自于对特定癌症的生物学和/或基因构成的了解。当这些因素被确定后，治疗就可以通过靶向癌症相关受体，从而使治疗具有肿瘤特异性，并减少非现场损伤的毒性。与其他实体肿瘤不同的是，G蛋白偶联受体(GPCRs)在调节胃肠道肿瘤的生长中起着关键作用。我们已经确定了胰腺癌上的一种受体，CCK-B受体，与正常组织相比，CCK-B受体在癌症中过度表达，并负责刺激生长。我们的研究团队已经证明，胃泌素通过GPCR以自分泌的方式调节其影响，从而刺激胰腺癌的生长。我们已经证明，如果胃泌素在受体上的相互作用能够被阻断，胰腺癌就会受到抑制。其中一个问题是开发一种有选择性地阻断这种受体的试剂，这种受体可能会被用于治疗。我们的实验室和其他实验室曾试图提出针对CCK-B受体胃泌素结合部位的单抗，但由于人和小鼠序列相似而未获成功。因此，我们采取了一种不同的策略，使用DNA适配子来靶向胰腺癌的CCK-B受体。DNA适配子是一种高度结构的寡核苷酸，它选择性地与受体靶标结合，亲和力与抗体相当或优于抗体，在外周血中比单抗更稳定，而且不具有免疫原性。我们合成了两个与CCK-B受体胞外N端的胃泌素结合部位相对应的多肽。使用一种称为指数富集型配体系统进化(SELEX)的技术，我们鉴定了与CCK-B受体的外部胃泌素结合位点结合的特定DNA适配子。我们假设CCK-B受体的选择性DNA适配子可以特异性地靶向胰腺癌，并通过阻断胃泌素的作用来抑制生长。为了验证这一假说，我们提出了以下目标：1)用体外培养的胰腺癌细胞鉴定一组适配子的结合亲和力(Kd)和IC50，以选择亲和力最高的适配子进行生长研究。2)研究适配子对胰腺癌体外培养和荷瘤小鼠生长的抑制作用。CCK-B靶向适配子在细胞凋亡中的作用也将被研究。我们研究的长期目标是提高胰腺癌患者的治疗水平和存活率。

项目成果

Preferential uptake of antibody targeted calcium phosphosilicate nanoparticles by metastatic triple negative breast cancer cells in co-cultures of human metastatic breast cancer cells plus bone osteoblasts.

• DOI: 10.1016/j.nano.2021.102383
• 发表时间: 2021-06
• 期刊: Nanomedicine : nanotechnology, biology, and medicine
• 作者: Bussard KM;Gigliotti CM;Adair BM;Snyder JM;Gigliotti NT;Loc WS;Wilczynski ZR;Liu ZK;Meisel K;Zemanek C;Mastro AM;Shupp AB;McGovern C;Matters GL;Adair JH
• 通讯作者: Adair JH