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.

项目摘要/摘要癌症免疫疗法的最新进展为三重患者提供了有希望的治疗选择阴性乳腺癌（TNBC）。尽管总体上成功治疗了这些恶性肿瘤，但免疫治疗性AP- 企业面临许多独特的挑战：（1）由于靶向副作用而引起的剂量限制，（2）加性毒性联合疗法的（3）和乳腺癌的免疫原性相对较低。为了克服这些限制，该提案旨在设计细菌的益生菌菌株，以选择性地定居于乳腺癌的肿瘤起源和局部释放治疗学。最终目标是创建可有效局部的诊所就绪菌株在维持患者的安全性的同时，ize和释放高剂量的治疗剂。为此，随附的项目将着重于工程基因编码的封装系统改善用于乳腺癌治疗的治疗细菌的静脉注射（IV）。在以前的研究中，我们有发现单次注射益生菌菌株E. Coli Nissle 1917（ECN）可以定居多个原发性和乳房起源的转移性肿瘤，与在此类器官中患有转移性疾病的TNBC患者有关作为肝，脑，骨骼和肺部。但是，在静脉注射遗传衰减细菌的临床试验中，不到15％的患者表现出有效的肿瘤定植，尽管安全给药剂量是著名的。使用合成生物学方法，我们先前设计了ECN来瞬时胶囊细菌表面上的多糖（称为ICAP），从而增加小鼠中最大耐受性的剂量 10倍。在第一个目标中，我们将基因组合该系统并将其与所使用的遗传衰减结合在我们建立的临床试验和其他安全系统中。我们将从益生菌中去除抗生素标记应变以使未来的翻译。由于对我们的ECN-ICAP的体液抗体反应将在人类，他们将限制系统产生多次注射的功效。因此，我们下一个构造两个其他封装系统（多氨酸，软骨素），以实现不同菌株的顺序输送。这些方法将通过多个体外测定和小鼠模型进行表征和测试。总体而言，使用工程益生菌的方法比当前的治疗层具有多个优点 egies，包括：（1）肿瘤特异性疗法的产生，（2）细菌裂解，导致有效释放新型治疗和脂多糖（LPS）辅助剂，以及（3）组合的功效和安全性增强国家和多个封装系统。这项工作旨在克服当前的免疫局限派，通过提供靶向车辆来局部提供刺激抗肿瘤免疫力的治疗剂，而预先预先排气系统性毒性并减轻与免疫相关的不良反应。