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: 8827374
• 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 2016-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8827374
• 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; Relative (related person); 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; 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、高度减毒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比其他组织高至少1000倍。人体临床研究已经验证了静脉内施用减毒细菌突变体的安全性，建立了耐受的多剂量，并且已经显示在一些患者中发生肿瘤靶向。然而，没有观察到抗肿瘤活性，即使在沙门氏菌被证实已在其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化细胞凋亡（程序性细胞死亡）诱导细胞毒性蛋白，能够杀死肿瘤细胞来克服。特定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，DNA的化学合成，构建编码蛋白质的基因，这些蛋白质将杀死癌细胞。这些合成蛋白的组成是基于已知的癌症杀伤和癌症靶向蛋白组分的组合，当组合时，产生具有选择性杀死癌症能力的蛋白。还将进行细胞凋亡和细胞毒性多肽的新型双功能融合蛋白的初步研究，以将其细胞靶向、亚细胞转运和细胞内分子靶向的联合收割机方面作为赋予增强的效力以及肿瘤特异性的手段，并为新的资助申请产生初步数据。 这里描述的细菌是高度安全的，适合学生在实验室工作。学生从PI教授的微生物学课程中招募，并整合了他对肿瘤靶向沙门氏菌的研究。学生也从研究生院入学计划，并在CSUN的MARC和RISE计划招募。虽然PI在CSUN只有一年半的时间，但他已经为11名学生提供了基于实验室的研究培训，其中包括3名西班牙裔，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。该项目预计将在两至三年内招收两名攻读硕士学位的研究生，在资助期间总共招收四名研究生，每年招收一至两名定向研究本科生;在资助期间，最多可招收8名学生，每学期可招收1至2名学生参加生物学研究课程，在资助期间，共有14至28名学生。