Programmable encapsulation systems to improve delivery of therapeutic bacteria

可编程封装系统可改善治疗性细菌的递送

基本信息

批准号：
10639259
负责人：
Nicholas Arpaia
金额：
$ 61.39万
依托单位：
COLUMBIA UNIV NEW YORK MORNINGSIDE
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10639259
关键词：
Adjuvant Adverse effects Antibiotics Antibody Response Attenuated Automobile Driving Bacteria Biodistribution Biological Assay Blood Body Weight Bone Tissue Brain Breast Cancer Model Breast Cancer Patient Breast Cancer therapy Cells Chondroitin Clinical Trials Colorectal Cancer Combined Modality Therapy Cytolysis Data Disease Dose Drops Encapsulated Engineered Probiotics Engineering Environment Escherichia coli Face Flagella Frequencies Future Genes Genetic Genetic Engineering Genomics Goals Human Immune Immune Evasion Immunologic Surveillance Immunotherapeutic agent Immunotherapy In Vitro In complete remission Individual Injections Intelligence Intravenous Knock-out Lipid A Lipopolysaccharides Liver Lung Malignant Neoplasms Mammary Neoplasms Maximum Tolerated Dose Measures Medicine Methods Modification Mus Mutation Neoplasm Metastasis Oral Organ Patients Polysaccharides Polysialic Acid Primary Neoplasm Probiotics Production Reporter Reporting Safety Serratia Soft tissue sarcoma Specificity Streptococcus Surface System Testing Therapeutic Toxic effect Toxin Translations Treatment Efficacy Tumor Burden Tumor Immunity Tumor Promotion Work bone cancer immunotherapy cancer therapy capsule clinic ready clinical candidate clinically relevant cytokine immunogenicity improved in vitro Assay in vivo intravenous administration intravenous injection malignant breast neoplasm microbial mouse model novel novel therapeutics patient safety preclinical trial prevent research clinical testing response safety assessment safety engineering safety testing side effect success synthetic biology systemic toxicity triple-negative invasive breast carcinoma tumor tumor hypoxia

项目摘要

PROJECT SUMMARY/ABSTRACT Recent advances in cancer immunotherapy have provided promising treatment options for patients with triple- negative breast cancer (TNBC). Despite overall success in treating these malignancies, immunotherapeutic ap- proaches face a number of unique challenges: (1) dose limitation due to off-target side effects, (2) additive toxicity of combination therapies, (3) and relatively low immunogenicity of breast cancer. To overcome these limitations, this proposal seeks to engineer probiotic strains of bacteria that selectively colonize tumors of breast-cancer origin and locally release therapeutics. The ultimate goal is to create clinic-ready strains that will efficiently local- ize and release high-doses of therapeutics, while maintaining safety for patients. To do so, the accompanying project will focus on engineering genetically encoded encapsulation systems to improve intravenous (IV) delivery of therapeutic bacteria for breast cancer therapy. In previous studies, we have found that a single injection of a probiotic strain E. coli Nissle 1917 (EcN) can colonize multiple primary and metastatic tumors of breast origin, relevant for TNBC patients that have metastatic disease across organs such as the liver, brain, bone, and lung. However, in clinical trials with IV injection of genetically-attenuated bacteria, less than 15% of patients demonstrated efficient colonization of tumors, although safe administration doses were noted. Using synthetic biology approaches, we previously engineered EcN for transient induction of capsule polysaccharides on the bacteria surface (termed iCAP), which increases maximally-tolerated doses in mice by 10-fold. In the first aim, we will genomically integrate this system and combine it with genetic attenuations used in clinical trials, and other safety systems we have built. We will remove antibiotic markers from the probiotic strain for future translation as well. Since humoral antibody responses to our EcN-iCAP will be generated in humans, they will limit the efficacy of the system to generate multiple injections. Thus, we will next construct two other encapsulation systems (polysialic acid, chondroitin), to enable sequential delivery of distinct strains. These approaches will be characterized and tested through multiple in vitro assays and in mouse models. Altogether the approach of using engineered probiotics has several advantages over current therapeutic strat- egies, including: (1) tumor-specific production of therapeutics, (2) bacteria lysis that leads to effective release of novel therapeutics and lipopolysaccharides (LPS) adjuvant, and (3) enhanced efficacy and safety from combi- nations and multiple encapsulation systems. This work seeks to overcome current limitations of immunothera- pies, by providing a targeted vehicle to locally deliver therapeutics that stimulate antitumor immunity while pre- venting systemic toxicity and mitigating immune-related adverse effects.

项目概要/摘要癌症免疫疗法的最新进展为三重癌症患者提供了有希望的治疗选择阴性乳腺癌（TNBC）。尽管在治疗这些恶性肿瘤方面取得了总体成功，但免疫治疗应用 Proaches面临着许多独特的挑战：（1）由于脱靶副作用导致的剂量限制，（2）附加毒性（3）乳腺癌的免疫原性相对较低。为了克服这些限制，该提案旨在设计能够选择性定植乳腺癌肿瘤的益生菌菌株起源和局部释放疗法。最终目标是创造可用于临床的菌株，以有效地本地化释放并释放高剂量的治疗药物，同时保证患者的安全。为此，随附的项目将重点关注基因编码封装系统的工程设计，以改善乳腺癌治疗细菌的静脉 (IV) 输送。在之前的研究中，我们有发现单次注射益生菌菌株 E. coli Nissle 1917 (EcN) 可以定植多个原代和乳腺起源的转移性肿瘤，与患有跨器官转移性疾病的 TNBC 患者相关，例如如肝脏、大脑、骨骼和肺。然而，在静脉注射基因减毒细菌的临床试验中，尽管安全给药剂量有限，但只有不到 15% 的患者表现出有效的肿瘤定植著名的。使用合成生物学方法，我们之前设计了用于瞬时诱导胶囊的 EcN 细菌表面的多糖（称为 iCAP），通过增加小鼠的最大耐受剂量 10倍。第一个目标是，我们将从基因组角度整合该系统，并将其与所使用的遗传衰减相结合在临床试验以及我们建立的其他安全系统中。我们将从益生菌中去除抗生素标记也为以后的翻译带来压力。由于对我们的 EcN-iCAP 的体液抗体反应将在人类，他们会限制系统产生多次注射的功效。因此，我们接下来将构建两个其他封装系统（聚唾液酸、软骨素），以实现不同菌株的顺序递送。这些将通过多种体外测定和小鼠模型对方法进行表征和测试。总的来说，使用工程益生菌的方法比目前的治疗策略有几个优点例如，包括：（1）肿瘤特异性治疗药物的产生，（2）细菌裂解导致有效释放新型疗法和脂多糖（LPS）佐剂，以及（3）增强组合的功效和安全性国家和多种封装系统。这项工作旨在克服目前免疫疗法的局限性馅饼，通过提供有针对性的载体来局部提供刺激抗肿瘤免疫力的治疗，同时预排出全身毒性并减轻免疫相关的不良反应。