Tumor-targeting Salmonella expressing apoptosis-inducing cytotoxic proteins

表达细胞凋亡诱导细胞毒性蛋白的肿瘤靶向沙门氏菌

• 批准号: 9026621
• 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 2018-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9026621
• 关键词: Admission activity; Algorithms; Antineoplastic Agents; Apoptosis; Attenuated; Bacteria; Biology; Breast Carcinoma; Breast Epithelial Cells; C-terminal; CCL7 gene; Cancerous; Cell Cycle Arrest; Cells; Chimera organism; Chimeric Proteins; Clinical Research; Clinical Trials; Combination Drug Therapy; Combined Modality Therapy; Cultured Tumor Cells; Cytotoxin; DNA chemical synthesis; Data; Disease; Distant; Dose; Drug Delivery Systems; Educational process of instructing; Engineering; Enrollment; Epitopes; Filipino; Funding; Future; Generations; Genes; Genetic Engineering; Grant; Health; Hispanics; Human; In Vitro; Individual; Injection of therapeutic agent; Intracellular Transport; Koreans; Laboratories; Ligands; M cell; MCF7 cell; Malignant Epithelial Cell; Malignant Neoplasms; Master' s Degree; Microbiology; Molecular Target; Normal Cell; Normal tissue morphology; Nuclear Localization Signal; Patients; Peptides; Production; Proteins; Recruitment Activity; Research; Research Training; Safety; Salmonella; Schools; Site; Solid Neoplasm; Specificity; Students; System; Testing; Therapeutic; Therapeutic Effect; Time; Tissues; Toxic effect; Tumor Cell Line; Viral Proteins; Work; attenuation; base; cancer cell; cancer therapy; cell type; cost; cytolethal distending toxin; cytotoxic; cytotoxicity; design; graduate student; in vivo; indexing; killings; mutant; neoplastic cell; novel; polypeptide; programs; research study; subcellular targeting; synthetic biology; synthetic construct; synthetic protein; therapeutic protein; tumor; tumor specificity; vector

DESCRIPTION (provided by applicant): 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 bacterial mutants, 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 which 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 apoptosis (programmed cell death) -inducing cytotoxic proteins that are able to kill tumor cells. The Specific Aims are designed to test individual proteins and protein combinations in order to develop tumor-targeted Salmonella vectors with enhanced antitumor activity. The project utilizes synthetic biology, the chemical synthesis of DNA, to construct genes encoding proteins that will kill cancer cells. The composition of these synthetic proteins is based on combinations of known cancer-killing and cancer-targeting protein components, which, when combined, result in proteins with the selective ability to kill cancer. A preliminary study of novel bifunctional fusios of the apoptosis and cytotoxic polypeptides will also be conducted in order to combine aspects of their cell targeting, subcellular transport and intracellular molecular targets as a means to confer enhanced potency together with tumor specificity, and to generate preliminary data for a new grant submission. The bacteria described here are highly safe and suitable for students to work on in the lab. Students are recruited from the microbiology courses the PI teaches and integrates his research on tumor-targeted Salmonella. Students are also recruited from the Graduate School Admission Program, and the MARC and RISE programs at CSUN. Although the PI has only been at CSUN for 1.5 years, he has already provided laboratory-based research training for 11 students, including 3 Hispanics, 2 Armenians, 2 Indians, 1 Filipino, 1 Thai, and 1 Korean. The project is expected to involve two graduate students seeking their Masters Degree over a two to three year period, with a total of up to four graduate students during the funding period, and one to two directed research undergraduates per year; up to eight during the course of the funding period, as well as one to two students per semester enrolled in the Biology research course, for a total of between 14 and 28 students during the funding period.

描述（由申请人提供）：肿瘤靶向细菌与其他癌症药物传递系统相比具有许多优势，包括从远处接种部位靶向多个肿瘤，选择性肿瘤内复制，高度衰减和安全性，以及直接在肿瘤内表达抗癌蛋白的能力。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其他组织的1000倍。人体临床研究已经证实了静脉注射减毒细菌突变体的安全性，建立了可耐受的多剂量，并表明在一些患者中发生了肿瘤靶向。然而，没有观察到抗肿瘤活性，即使在沙门氏菌已被证实在其肿瘤上定植的患者中也是如此。我们假设在人类临床试验中缺乏抗肿瘤功效可以通过工程细胞凋亡（程序性细胞死亡）来克服，这种细胞凋亡诱导细胞毒性蛋白能够杀死肿瘤细胞。特异性目标旨在测试单个蛋白质和蛋白质组合，以开发具有增强抗肿瘤活性的肿瘤靶向沙门氏菌载体。该项目利用合成生物学，即DNA的化学合成，来构建编码蛋白质的基因，从而杀死癌细胞。这些合成蛋白质的组成是基于已知的癌症杀伤和癌症靶向蛋白质成分的组合，当它们结合在一起时，就会产生具有选择性杀死癌症能力的蛋白质。一项关于细胞凋亡和细胞毒性多肽新型双功能融合的初步研究也将进行，以便将它们的细胞靶向、亚细胞转运和细胞内分子靶点结合起来，作为一种赋予增强效力和肿瘤特异性的手段，并为新的拨款申请提供初步数据。这里描述的细菌是高度安全的，适合学生在实验室工作。学生是从PI教授的微生物学课程中招募的，并整合了他对肿瘤靶向沙门氏菌的研究。学生也从CSUN的研究生入学计划，MARC和RISE计划中招募。虽然PI只在CSUN工作了一年半，但他已经为11名学生提供了实验室研究培训，其中包括3名西班牙人，2名亚美尼亚人，2名印度人，1名菲律宾人，1名泰国人和1名韩国人。该项目预计将有两名研究生在两至三年的时间内攻读硕士学位，在资助期内总共有四名研究生，每年有一至两名本科生进行定向研究；资助期内最多招收8名学生，每学期招收1至2名学生参加生物学研究课程，资助期内总共招收14至28名学生。