TUMOR-TARGETING SALMONELLA EXPRESSING TUMOR-SELECTIVE CYTOTOXIC PROTEINS IN COMBINATION WITH PROTEASE INHIBITORS

表达肿瘤选择性细胞毒性蛋白的肿瘤靶向沙门氏菌与蛋白酶抑制剂的组合

• 批准号: 10321211
• 负责人: DAVID G BERMUDES
• 金额: $ 10.88万
• 依托单位: CALIFORNIA STATE UNIVERSITY NORTHRIDGE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-10 至 2024-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10321211
• 关键词: Animals; Antineoplastic Agents; Apoptosis; Attenuated; Bacteria; Biodistribution; Breast Cancer Model; Cancer Model; Cancerous; Chimera organism; Clinical Research; Clinical Trials; Data; Development; Disease; Distant; Dose; Drug Delivery Systems; Effectiveness; Engineering; Epidermal Growth Factor Receptor; Funding; Generations; Goals; Human; In Vitro; Individual; Intravenous; Lead; Malignant Neoplasms; Metalloproteases; Modeling; Mus; Pathogenicity; Patients; Peptide Hydrolases; Process; Protease Inhibitor; Proteins; Proteolysis; Pseudomonas aeruginosa toxA protein; Research; Safety; Salmonella; Site; Solid Neoplasm; Sunflowers; Targeted Toxins; Testing; Therapeutic Agents; Time; Tissues; Toxic effect; Toxin; Trypsin Inhibitors; Xenograft procedure; aggressive breast cancer; anti-cancer; anticancer activity; antitumor effect; attenuation; base; cancer cell; cancer therapy; cytotoxic; design; human model; improved; in vivo; inhibitor; matriptase; mouse model; mutant; neoplastic cell; optical imaging; prevent; protein degradation; therapeutic protein; translational study; triple-negative invasive breast carcinoma; tumor; tumor xenograft; vector; whole body imaging

PROJECT SUMMARY/ABSTRACT Tumor-targeting bacteria offer a number of advantages over other cancer drug delivery systems, including targeting of multiple tumors from a distant inoculation site, selective intratumoral replication, a high degree of attenuation and safety, and the ability to express anti-cancer proteins directly within the tumor. Remarkably, attenuated Salmonella localize within solid tumors at levels at least 1000 times greater than other tissues. Human clinical studies have validated the safety of intravenously administered attenuated Salmonella mutant VNP20009, established tolerated multiple doses, and have shown that tumor targeting occurs in some patients. However, no anti-tumor activity was observed, even in patients in whom the Salmonella were verified to have colonized their tumors. We hypothesize that the lack of antitumor efficacy in the human clinical trials can be overcome by engineering antitumor apoptosis (programmed cell death) -inducing cytotoxic proteins that are able to selectively kill tumor cells, and that the ability of these proteins to kill cancer cells can be enhanced by blocking their degradation by tumor proteases through co-expression of protease inhibitors. The specific aims are designed to test the tumor-selective toxin generated during the initial SC3 funding period in murine models of cancer. The specific aims are also directed toward generation and analysis of individual protease inhibitor combinations expressed by VNP20009 in order to prevent proteolytic degradation of the therapeutic protein and develop tumor-targeted Salmonella vectors with enhanced antitumor activity. VNP20009 expressing a tumor-cell targeted toxin will be analyzed alone and in combination with one or more protease inhibitors using optical imaging of the bacterial interaction with murine tumor models of highly aggressive breast cancer. The results are expected to indicate the effectiveness of the targeted toxin and the effect(s) of the protease inhibitors alone and in combination with the toxin. These results have the potential to improve VNP20009 without increasing its toxicity, and therefore could lead to translational studies in humans. .

项目总结/摘要 肿瘤靶向细菌提供了许多优于其他癌症药物递送系统的优点， 包括从远处接种部位靶向多个肿瘤，选择性肿瘤内复制， 高度的减毒和安全性，以及直接在肿瘤细胞内表达抗癌蛋白的能力。 肿瘤值得注意的是，减毒沙门氏菌在实体瘤中的定位水平至少是 高于其他组织。人体临床研究已经证实了静脉注射 给予减毒沙门氏菌突变体VNP 20009，建立耐受多剂量，并具有 显示肿瘤靶向发生在一些患者中。然而，没有观察到抗肿瘤活性， 即使在那些沙门氏菌被证实已经在肿瘤中定植的患者中也是如此。我们假设 在人体临床试验中缺乏抗肿瘤疗效可以通过工程抗肿瘤 能够选择性杀死肿瘤的凋亡（程序性细胞死亡）诱导细胞毒性蛋白 这些蛋白质杀死癌细胞的能力可以通过阻断它们的作用来增强。 通过蛋白酶抑制剂的共表达被肿瘤蛋白酶降解。具体目标是 设计用于在小鼠中测试最初SC 3资助期间产生的肿瘤选择性毒素 癌症模型。具体的目标也是针对个人的生成和分析 VNP 20009表达的蛋白酶抑制剂组合，以防止 治疗性蛋白质和开发具有增强的抗肿瘤作用的肿瘤靶向沙门氏菌载体 活动将单独和组合分析表达肿瘤细胞靶向毒素的VNP 20009 使用细菌与鼠肿瘤相互作用的光学成像， 高度侵袭性乳腺癌的模型。预计结果将表明该方法的有效性 靶向毒素和蛋白酶抑制剂单独和与毒素组合的作用。 这些结果有可能改善VNP 20009而不增加其毒性，因此 可以用于人类的转化研究。 .

项目成果

EGFR-targeted Chimeras of Pseudomonas ToxA released into the extracellular milieu by attenuated Salmonella selectively kill tumor cells.

• DOI: 10.1002/bit.26026
• 发表时间: 2016-12
• 期刊: BIOTECHNOLOGY AND BIOENGINEERING
• 影响因子: 3.8
• 作者: Quintero, David;Carrafa, Jamie;Vincent, Lena;Bermudes, David
• 通讯作者: Bermudes, David

A culture-based method for determining the production of secreted protease inhibitors.

一种基于培养的方法，用于确定分泌型蛋白酶抑制剂的产生。

• DOI: 10.1016/j.mimet.2014.02.019
• 发表时间: 2014
• 期刊: Journal of microbiological methods
• 影响因子: 2.2
• 作者: Quintero,David;Bermudes,David
• 通讯作者: Bermudes,David

Accumulation of single-stranded DNA in Escherichia coli carrying the colicin plasmid pColE3-CA38.

• DOI: 10.1016/j.plasmid.2014.11.001
• 发表时间: 2015-01
• 期刊: Plasmid
• 影响因子: 2.6
• 作者: Morales M;Attai H;Troy K;Bermudes D
• 通讯作者: Bermudes D

Co-Expression of a Chimeric Protease Inhibitor Secreted by a Tumor-Targeted Salmonella Protects Therapeutic Proteins from Proteolytic Degradation.

靶向肿瘤沙门氏菌分泌的嵌合蛋白酶抑制剂的共表达可保护治疗性蛋白质免遭蛋白水解降解。

• DOI: 10.4014/jmb.1807.08036
• 发表时间: 2018
• 期刊: Journal of microbiology and biotechnology
• 影响因子: 2.8
• 作者: Quintero,David;Carrafa,Jamie;Vincent,Lena;Lee,HeeJong;Wohlschlegel,James;Bermudes,David
• 通讯作者: Bermudes,David