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）：癌症疗法的进展均来自了解特定癌症的生物学和或基因组成。当鉴定出这些因素时，可以通过靶向与癌症相关的受体来“个性化”，并通过肿瘤特异性“个性化”，而异地损伤的毒性较小。与其他实体瘤不同，G蛋白偶联受体（GPCR）已被证明在胃肠道癌的生长中起着至关重要的作用。我们已经确定了胰腺癌的受体CCK-B受体，该受体与正常组织相比在癌症中过表达，并负责生长刺激。我们的研究小组表明，胃蛋白通过这种GPCR以自分泌方式介导其作用来刺激胰腺癌的生长。我们已经表明，如果可以阻断胃癌在受体上的相互作用，则会抑制胰腺癌。一个问题是开发一种可以选择性地阻断可能在治疗方法上使用的受体的药物。我们的实验室和其他实验室试图对CCK-B受体的胃蛋白结合位点提出单克隆抗体，但由于人类和小鼠序列之间的类似同源性，因此没有成功。因此，我们使用DNA适体在胰腺癌中靶向CCK-B受体的策略。 DNA适体 - 与单克隆抗体相比，在外周血中选择性或比抗体相当或更好的亲和力的受体靶标结合的高度结构化的寡核苷酸，并且不是免疫原性。我们合成了对应于CCK-B受体的细胞外N末端上胃蛋白结合位点的两种肽。使用指数富集（SELEX）称为配体的系统演化的技术，我们确定了与CCK-B受体的外部胃蛋白结合位点结合的特定DNA适体。我们假设对CCK-B受体的选择性DNA适体可以特异性地靶向胰腺癌并通过阻止胃蛋白的作用来抑制生长。为了检验这一假设，我们提出以下目的：1）表征我们使用胰腺癌细胞在体外使用胰腺癌细胞确定的适体小组的结合亲和力（KD）和IC50，以选择具有生长研究最大亲和力的适体。 2）研究适体抑制胰腺癌生长在培养物和伴有原位胰腺癌的小鼠中的功效。 CCK-B靶向适体在凋亡中的作用也将进行研究。我们研究的长期目标是改善胰腺癌患者的治疗和存活。

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