3D Printed Engineered Living Materials for Drug Delivery

用于药物输送的 3D 打印工程活性材料

基本信息

批准号：
10602505
负责人：
Alshakim Nelson
金额：
$ 18.17万
依托单位：
UNIVERSITY OF WASHINGTON
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-05 至 2024-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10602505
关键词：
3-Dimensional 3D Print Acrylates Affect Alkaloids American Anti-Inflammatory Agents Bacteria Berberine Biodegradation Biological Availability Biological Preservation Biological Process Biomedical Technology Caco-2 Cells Cells Clinical Colon Carcinoma Colorectal Cancer Crohn&apos s disease Cytoprotection Development Devices Disease Drug Delivery Systems Engineering Epithelium Escherichia coli Excision Formulation Gastrointestinal Diseases Goals Growth Human Microbiome Hydrogels Immobilization In Situ In Vitro Inflammation Inflammatory Inflammatory Bowel Diseases Intestinal Diseases Intestines Irritable Bowel Syndrome Malignant Neoplasms Medical Device Microbe Modeling Modulus Needles Outcome Patients Performance Pharmaceutical Preparations Plant Resins Polymers Production Proteins Public Health Publishing Research Stents System Technology Testing Therapeutic Therapeutic Agents Time Translations Treatment Efficacy Ulcerative Colitis Validation Variant Water Weight Work Yeasts aqueous copolymer cytotoxicity design digital models drug production effective therapy experimental study fabrication globular protein gut microbiome improved in vitro Model in vivo local drug delivery mechanical properties metabolic engineering microorganism monomer next generation polymerization prevent programs prototype response synthetic biology treatment duration tumor virtual

项目摘要

PROJECT SUMMARY Traditional drug delivery platforms are limited by the amount of drug that can be loaded into the delivery system. While drug loading capacity can reach 50% by weight for some polymeric systems, the time period over which the delivery of the therapeutic can be sustained is often limited. In addition, the therapeutic efficacy of a local drug delivery system is related to the local bioavailability of the active agent. Devices that bio-catalytically produce the therapeutic in situ could thus provide more effective local delivery of the active drug and improve therapeutic efficacy. The long-term vision for this program is to create the next-generation 3D printed in situ drug production and delivery devices, including drug-eluting stents, microneedles, and patches, based on engineered living materials (ELMs) for localized and sustained therapeutic delivery. ELMs are composites of microorganisms incorporated within a polymeric matrix, wherein the cells maintain their viability and can be metabolically engineered to produce a therapeutic compound. Despite the immense potential of ELMs for drug delivery applications, the primary challenges to the deployment of ELMs as drug-eluting stents or patches to treat intestinal diseases are that ELMs must (i) be processable into precise form factors (e.g., patient-specific stents) with the requisite mechanical properties, (ii) biodegrade into non-cytotoxic components at predetermined rates, and (iii) bio-catalytically produce and elute the therapeutic agent in situ for the lifetime of the device. The objective of this proposal is to develop 3D printable resins that afford biodegradable hydrogel constructs with a tunable stiffness, and to demonstrate the fabrication of a prototype ELM device for sustained delivery of a model compound. In Aim 1, we will create aqueous resins with non-pathogenic E. coli Nissle 1917 (EcN) that can be 3D printed into hydrogel constructs using a commercially available 3D printer. We will formulate aqueous resins comprised of soluble globular protein derivatives that can be co-polymerized with water-soluble acrylate monomers upon photo-initiated polymerization. The mechanical properties of the ELM hydrogels (stiffness, strength, and toughness) and rates of enzymatic degradation will be quantified for each resin formulation. In Aim 2, we will metabolically engineer non-pathogenic EcN to produce berberine as a model therapeutic compound. We will further evaluate the cytotoxicity and epithelial integrity of Caco-2 cells in the presence of 3D printed ELMs. As validation of these ELMs we will use an in vitro model to confirm the production and elution of berberine from the 3D printed ELM by evaluating the Caco-2 response to proinflammatory stimulation. We envision these 3D printed ELMs to be used as devices for local drug delivery in the treatment of malignancies or inflammatory diseases affecting the intestines. While our ELM platform is compatible with a broad array of microorganisms that include yeast and bacteria, we have chosen to focus on engineered variants of E. coli Nissle 1917, a commensal strain of bacteria common within the gut microbiome. Using a microorganism native to the human microbiome may facilitate translation of our platform to treat intestinal diseases.

项目概要传统的药物输送平台受到可装载到输送系统中的药物量的限制。虽然某些聚合物系统的载药量可以达到重量的 50%，但治疗的持续递送通常是有限的。另外，局部治疗的效果药物递送系统与活性剂的局部生物利用度有关。生物催化装置原位产生治疗剂因此可以提供活性药物更有效的局部递送并改善治疗功效。该项目的长期愿景是创造下一代 3D 打印原位药物基于工程设计的生产和输送设备，包括药物洗脱支架、微针和贴片用于局部和持续治疗传递的活性材料（ELM）。 ELM 是微生物的复合材料掺入聚合物基质中，其中细胞保持其活力并可以代谢设计用于生产治疗化合物。尽管 ELM 在药物输送方面具有巨大潜力应用，将 ELM 部署为药物洗脱支架或贴片来治疗的主要挑战肠道疾病的特点是 ELM 必须 (i) 可加工成精确的形状因素（例如患者特异性支架）具有必要的机械性能，（ii）以预定的速率生物降解为非细胞毒性成分， (iii)在装置的使用寿命期间原位生物催化地产生和洗脱治疗剂。目标该提案的目的是开发可 3D 打印的树脂，提供可生物降解的水凝胶结构，并具有可调的刚度，并演示用于持续交付模型的原型 ELM 设备的制造化合物。在目标 1 中，我们将使用非致病性大肠杆菌 Nissle 1917 (EcN) 制造水性树脂，该树脂可以使用市售 3D 打印机 3D 打印成水凝胶结构。我们将配制水性树脂由可溶性球状蛋白衍生物组成，可与水溶性丙烯酸酯共聚光引发聚合时的单体。 ELM 水凝胶的机械性能（刚度、强度和韧性）和酶降解速率将针对每种树脂配方进行量化。瞄准 2，我们将通过代谢工程非致病性 EcN 来生产小檗碱作为模型治疗化合物。我们将进一步评估 Caco-2 细胞在 3D 打印的情况下的细胞毒性和上皮完整性 ELM。作为对这些 ELM 的验证，我们将使用体外模型来确认小檗碱的产生和洗脱通过评估 Caco-2 对促炎刺激的反应，从 3D 打印的 ELM 中获得结果。我们设想这些 3D 打印 ELM 可用作治疗恶性肿瘤或炎症的局部药物输送装置影响肠道的疾病。我们的 ELM 平台与多种微生物兼容包括酵母和细菌，我们选择重点关注大肠杆菌 Nissle 1917 的工程变体，这是一种肠道微生物组中常见的细菌共生菌株。使用人类原生微生物微生物组可能有助于我们的平台转化为治疗肠道疾病。