Improved Tumor Targeting of Salmonella VNP20009 via Ice-llama Antibody Guidance

通过 Ice-llama 抗体指导改进沙门氏菌 VNP20009 的肿瘤靶向

• 批准号: 8637021
• 负责人: ANDREW HAYHURST
• 金额: $ 23.17万
• 依托单位: TEXAS BIOMEDICAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8637021
• 关键词: 3-Dimensional; Adverse effects; Animal Model; Antibiotics; Antibodies; Attenuated; Bacteria; Benchmarking; Binding; Bioluminescence; Breast Cancer Cell; Cancer Patient; Cancer cell line; Carcinoembryonic Antigen; Cause of Death; Cell Culture Techniques; Cell surface; Cells; Cessation of life; Colony-forming units; Diagnosis; Drug Delivery Systems; Engineering; Enzyme-Linked Immunosorbent Assay; Equilibrium; Fluorescence Microscopy; Gastrointestinal Neoplasms; Gene Fusion; Genetic; Goals; Growth; Heart Diseases; Human; Ice; Immunohistochemistry; In Situ; In Vitro; Link; Llama; Longitudinal Studies; MCF7 cell; Malignant Neoplasms; Measurement; Mediating; Membrane Proteins; Metastatic Melanoma; Modeling; Molecular; Monitor; Nanostructures; Necrosis; Organ; Outcome; Patients; Plasmids; Proteins; Radio; Resistance; Salmonella; Salmonella enterica; Solutions; Staging; Surface; System; Therapeutic; Time; Tissue Harvesting; Titrations; Toxic effect; Tumor Cell Invasion; Tumor Markers; Virus; Woman; Xenograft Model; antigen binding; base; design; improved; in vivo; luminescence; malignant breast neoplasm; mouse model; neoplastic cell; public health relevance; research study; tumor

DESCRIPTION (provided by applicant): ABSTRACT While Salmonella VNP 20009 (VNP) has shown promise in tumor targeting in animal models it is becoming apparent that regrowth of tumors is the outcome since the bacteria preferentially grow in the necrotic core and fail to reach the vital periphery. In human trials of metastatic melanoma patients, even targeting itself was very poor and no tumor regression was observed. We aim to improve the tumor targeting and tumor regressive ability of VNP by conferring upon it the successful recognition potential of antibody mediated tumor targeting. We had previously attempted to do this using the lpp-OmpA surface display platform and poorly soluble scFv antibodies but have since found out that the combination caused bacterial growth arrest and restricted the therapeutic potential of engineered VNP. We hypothesize that VNP displaying small and highly soluble anti- CEA single domain antibodies via the ice nucleation protein (INP) will show enhanced targeting of CEA positive tumors without inhibiting bacterial invasion or replication and will therefore be able to regress tumors more effectively than parental VNP. Our specific aims are to; 1, Engineer VNP to display anti-CEA sdAb without impeding replication, yet capable of binding immobilized CEA in vitro; 2, Demonstrate engineered VNP is capable of being internalized in MCF-7 cells and is also capable of homogenous targeting in three- dimensional spheroidal cell culture models; 3, Demonstrate improved in vivo tumor targeting of engineered VNP in a mouse model of breast cancer, elucidating intra-tumoral distribution and potential for complete tumor regression. The multifunctional capabilities of VNP including mobility and ability to carry large payloads are well balanced by the ability to visualize single bioluminescent cells and facile elimination of the bacteria by antibiotics. As such, the potential for VNP as a highly controllable yet versatile cancer therapeutic is enormous so long as we can improve targeting and regressive capacity.

摘要：虽然沙门氏菌VNP 20009 （VNP）在动物模型中显示出肿瘤靶向的前景，但由于细菌优先生长在坏死的核心并且无法到达，因此肿瘤的再生是显而易见的结果