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

基本信息

批准号：
8414763
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
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-10 至 2017-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8414763
关键词：
Admission activity Algorithms Antineoplastic Agents Apoptosis Attenuated Bacteria Biology Breast Carcinoma 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 Epithelial Cells Epitopes Filipino Funding Future Generations Genes Genetic Engineering Grant Hispanics Human In Vitro Individual Injection of therapeutic agent Intracellular Transport Koreans Laboratories Ligands M cell MCF7 cell Malignant Epithelial Cell Malignant Neoplasms Mammary gland Master&apos 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 public health relevance research study synthetic biology synthetic construct synthetic protein therapeutic protein tumor tumor specificity vector

项目摘要

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）：与其他癌症药物递送系统相比，肿瘤靶向细菌具有许多优势，包括从远处接种位点靶向多个肿瘤、选择性肿瘤内复制、高度减毒和安全性以及能够直接在肿瘤内表达抗癌蛋白。值得注意的是，减毒沙门氏菌在实体瘤内的定位水平至少比其他组织高 1000 倍。人体临床研究已经验证了静脉注射减毒细菌突变体的安全性，建立了耐受的多次剂量，并表明肿瘤靶向发生在一些患者身上。然而，即使在沙门氏菌被证实已在肿瘤中定植的患者中，也没有观察到抗肿瘤活性。我们假设，人类临床试验中抗肿瘤功效的缺乏可以通过设计细胞凋亡（程序性细胞死亡）来克服，诱导能够杀死肿瘤细胞的细胞毒性蛋白。具体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）来构建编码能够杀死癌细胞的蛋白质的基因。这些合成蛋白质的组成基于已知的抗癌和靶向癌症蛋白质成分的组合，当组合时，会产生具有选择性杀死癌症能力的蛋白质。还将对凋亡和细胞毒性多肽的新型双功能融合进行初步研究，以便将其细胞靶向、亚细胞转运和细胞内分子靶标的各个方面结合起来，作为赋予增强效力和肿瘤特异性的手段，并生成初步数据提交新的赠款。这里描述的细菌非常安全，适合学生在实验室研究。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。学生还从研究生院招生计划以及 CSUN 的 MARC 和 RISE 计划中招募。尽管PI在CSUN工作仅1.5年，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。该项目预计将涉及两名在两到三年内攻读硕士学位的研究生，资助期间总共最多四名研究生，每年还有一到两名定向研究型本科生；资助期内最多有 8 名学生，每学期有 1 至 2 名学生参加生物学研究课程，资助期内总共有 14 至 28 名学生。