Targeted tumoricidal bacteria

靶向杀肿瘤细菌

• 批准号: 7219081
• 负责人: MING ZHAO
• 金额: $ 14.98万
• 依托单位: ANTICANCER, INC.
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-21 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7219081
• 关键词: Amino Acids; Anaerobic Bacteria; Animals; Area; Bacteria; Color; Condition; Disseminated Malignant Neoplasm; Engineering; Future; Goals; Grant; Green Fluorescent Proteins; Human; Hypoxia; Image; Imagery; Imaging technology; Immune system; Immunocompetent; Infection; Label; Laboratories; Malignant - descriptor; Malignant Neoplasms; Mammary Neoplasms; Modeling; Mus; Nature; Necrosis; Normal tissue morphology; Nude Mice; Nutritional Requirements; Patients; Phase; Probability; Prostate; Proteins; Radiology Specialty; Salmonella typhimurium; Specificity; System; Testing; Tissues; Tumor Tissue; Variant; Virulent; cancer cell; cancer therapy; chemotherapeutic agent; clinical application; immunological status; killings; leucylarginine; microorganism; mouse model; mutant; red fluorescent protein; research study; response; tumor; whole body imaging

DESCRIPTION (provided by applicant): The goal of this application is to develop imageable tumor-targeting bacteria that can cure tumors without progressive infection of the host. Previous experiments by others employed anaerobic microorganisms for cancer therapy. Target specificity appeared largely due to the anaerobic requirements met principally in necrotic tumor areas. The resulting tumor killing was at best limited since anaerobic bacteria could not grow in viable tumor tissue. Therefore, more effective targeting is necessary, especially in the viable tumor tissue. Toward this goal, we have recently developed whole-body imaging systems that enable the visualization of green fluorescent protein (GFP)- and red fluorescent protein (RFP)- expressing tumors and bacteria (Nature Reviews Cancer 5, 796-806, 2005; Proc. Natl. Acad. Sci. USA 98, 9814-9818, 2001). With the help of the imaging technology, we have developed a unique tumor-targeting Salmonella typhimurium strain (Proc. Natl. Acad. Sci USA 102, 755-760, 2005). This strain is an auxotrophic but fully virulent variant of the facultative anaerobe Salmonella typhimurium, termed A1 that can grow under hypoxic or normoxic conditions. The A1 auxotrophic strain selectively grows in and destroys viable as well as necrotic malignant tissue but has little effect on normal tissue. The A1 bacteria eventually disappears from normal tissue even in immunodeficient nude mice. We rapidly selected this bacteria by labeling them with GFP and imaging their ability to target tumor labeled with RFP. This remarkable selectivity apparently reflects the imposed nutritional requirements that are apparently met only in the cancer cell milieu. S. typhimurium A1 is a double amino acid auxotroph that requires Leu and Arg. We have now demonstrated that human prostate and breast tumor mouse models have highly significant survival increases when treated with A1. The goal of this application is to expand the tumor types targeted by auxotrophic S. typhimurium and to determine if the host immune system enhances tumor kill by the bacteria. The specific aims are as follows: (1) Select additional multiple amino-acid auxotrophs of S. typhimurium to expand tumor-killing selectivity to additional tumor types, including patient tumor models; (2) Determine possible significance of host immunological status by comparing antitumor efficacy of selected S. typhimurium auxotrophs in nude-mouse and immunocompetent-mouse tumor models. In the Phase II grant, the effective antitumor bacterial strains will be further developed for eventual clinical application. Deletion mutants will be developed to reduce the probability of reversion of auxotrophs to wild-type. Determination of possible synergy of tumor targeting S. typhimurium auxotrophic strains and chemotherapeutic agents as well as radiology will also be tested in the Phase II application. Bacterial therapy for metastatic cancer is described. Genetically-altered bacteria that grow only in tumors and destroy them are being developed. Both bacteria and tumors are engineered to fluoresce different colors such that the targeting of the bacteria to the tumors can be visualized external to the mouse models being used. Future human trials of the tumor-killing bacteria can be held after the Phase I and Phase II grant periods are completed.

描述（由申请人提供）：本申请的目标是开发可成像的肿瘤靶向细菌，其可以治愈肿瘤而不会对宿主进行性感染。其他人以前的实验使用厌氧微生物进行癌症治疗。靶特异性主要是由于坏死肿瘤区域主要满足厌氧要求。由于厌氧菌不能在活的肿瘤组织中生长，因此产生的肿瘤杀伤最多是有限的。因此，更有效的靶向是必要的，特别是在活的肿瘤组织中。为了这个目标，我们最近开发了全身成像系统，其使得能够可视化表达绿色荧光蛋白（GFP）和红色荧光蛋白（RFP）的肿瘤和细菌（Nature Reviews Cancer 5，796-806，2005; Proc. Natl. Acad. Sci. USA 98，9814-9818，2001）。在成像技术的帮助下，我们已经开发了一种独特的肿瘤靶向鼠伤寒沙门氏菌菌株（Proc. Natl. Acad. Sci USA 102，755-760，2005）。该菌株是兼性厌氧菌鼠伤寒沙门氏菌（称为A1）的营养缺陷型但完全毒性变体，可在低氧或常氧条件下生长。A1营养缺陷型菌株选择性地在存活和坏死的恶性组织中生长并破坏存活和坏死的恶性组织，但对正常组织影响不大。即使在免疫缺陷的裸鼠中，A1细菌最终也会从正常组织中消失。我们通过用GFP标记它们并成像它们靶向RFP标记的肿瘤的能力来快速选择这种细菌。这种显著的选择性显然反映了明显仅在癌细胞环境中满足的强加的营养需求。S.鼠伤寒A1是需要Leu和Arg的双氨基酸营养缺陷型。我们现在已经证明，当用A1治疗时，人前列腺和乳腺肿瘤小鼠模型具有非常显著的存活率增加。本申请的目的是扩大营养缺陷型S.并确定宿主免疫系统是否增强细菌对肿瘤的杀伤。具体目标如下：（1）筛选出附加的多氨基酸营养缺陷型菌株。鼠伤寒沙门氏菌，以扩大肿瘤杀伤选择性的其他肿瘤类型，包括患者的肿瘤模型;（2）确定宿主免疫状态的可能意义，通过比较选定的S.裸鼠和免疫活性小鼠肿瘤模型中的鼠伤寒沙门氏菌营养缺陷型。在II期资助中，将进一步开发有效的抗肿瘤菌株，以供最终临床应用。将开发缺失突变体以降低营养缺陷型回复为野生型的可能性。确定肿瘤靶向S.鼠伤寒营养缺陷型菌株和化疗剂以及放射学也将在II期申请中进行测试。描述了用于转移性癌症的细菌疗法。目前正在开发一种只在肿瘤中生长并能破坏肿瘤的转基因细菌。细菌和肿瘤都被设计成发出不同颜色的荧光，这样细菌对肿瘤的靶向就可以在所使用的小鼠模型外部可视化。肿瘤杀伤细菌的未来人体试验可以在第一阶段和第二阶段资助期完成后进行。

项目成果

A color-coded imaging model of the interaction of αv integrin-GFP expressed in osteosarcoma cells and RFP expressing blood vessels in Gelfoam® vascularized in vivo.

骨肉瘤细胞中表达的αv整合素-GFP与体内血管化的Gelfoam®中表达血管的RFP相互作用的颜色编码成像模型。

• 发表时间: 2013
• 期刊: Anticancer research
• 影响因子: 2
• 作者: Uehara,Fuminari;Tome,Yasunori;Yano,Shuya;Miwa,Shinji;Mii,Sumiyuki;Hiroshima,Yukihiko;Bouvet,Michael;Maehara,Hiroki;Kanaya,Fuminori;Hoffman,RobertM
• 通讯作者: Hoffman,RobertM

Dynamic color-coded fluorescence imaging of the cell-cycle phase, mitosis, and apoptosis demonstrates how caffeine modulates cisplatinum efficacy.

细胞周期阶段、有丝分裂和细胞凋亡的动态颜色编码荧光成像展示了咖啡因如何调节顺铂功效。

• DOI: 10.1002/jcb.24593
• 发表时间: 2013
• 期刊: Journal of cellular biochemistry
• 影响因子: 4
• 作者: Miwa,Shinji;Yano,Shuya;Tome,Yasunori;Sugimoto,Naotoshi;Hiroshima,Yukihiko;Uehara,Fuminari;Mii,Sumiyuki;Kimura,Hiroaki;Hayashi,Katsuhiro;Efimova,ElenaV;Fujiwara,Toshiyoshi;Tsuchiya,Hiroyuki;Hoffman,RobertM
• 通讯作者: Hoffman,RobertM

Primer dosing of S. typhimurium A1-R potentiates tumor-targeting and efficacy in immunocompetent mice.

鼠伤寒沙门氏菌 A1-R 的引物给药可增强免疫活性小鼠的肿瘤靶向性和功效。

• 发表时间: 2013
• 期刊: Anticancer research
• 影响因子: 2
• 作者: Tome,Yasunori;Zhang,Yong;Momiyama,Masashi;Maehara,Hiroki;Kanaya,Fuminori;Tomita,Katsuro;Tsuchiya,Hiroyuki;Bouvet,Michael;Hoffman,RobertM;Zhao,Ming
• 通讯作者: Zhao,Ming

Salmonella typhimurium A1-R tumor targeting in immunocompetent mice is enhanced by a traditional Chinese medicine herbal mixture.

• 发表时间: 2013-05
• 期刊: Anticancer research
• 影响因子: 2
• 作者: Yong Zhang;N. Zhang;S. Su;R. Hoffman;Ming Zhao

Single cell time-lapse imaging of focus formation by the DNA damage-response protein 53BP1 after UVC irradiation of human pancreatic cancer cells.

人胰腺癌细胞 UVC 照射后 DNA 损伤反应蛋白 53BP1 形成焦点的单细胞延时成像。

• 发表时间: 2013
• 期刊: Anticancer research
• 影响因子: 2
• 作者: Miwa,Shinji;Tome,Yasunori;Yano,Shuya;Hiroshima,Yukihiko;Uehara,Fuminari;Mii,Sumiyuki;Kimura,Hiroaki;Hayashi,Katsuhiro;Tsuchiya,Hiroyuki;Bouvet,Michael;Efimova,ElenaV;Hoffman,RobertM
• 通讯作者: Hoffman,RobertM