3D Printed Engineered Living Materials for Drug Delivery

用于药物输送的 3D 打印工程活性材料

• 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
• 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' 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

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.

项目摘要 传统的药物输送平台受到可以加载到输送系统的药物量的限制。 对于某些聚合系统，药物加载能力可以达到50％的重量，但 可以维持治疗的交付通常受到限制。另外，局部的治疗功效 药物输送系统与活性剂的局部生物利用度有关。生物催化的设备 因此，产生原位治疗性可以提供活性药物的更有效的局部输送 治疗功效。该程序的长期愿景是创建下一代印刷的原位药物 基于工程的生产和递送设备，包括洗脱支架，微针和补丁 生物材料（ELMS）用于局部和持续的治疗性分娩。榆树是微生物的复合材料 掺入聚合物基质中，其中细胞保持其生存力，可以代谢 设计用于生产治疗化合物。尽管ELM具有巨大的药物输送潜力 申请，将榆树作为药物洗脱支架或补丁部署的主要挑战 肠道疾病是ELMS必须（i）可处理成精确的形式（例如，患者特定的支架） 具有必要的机械性能，（ii）以预定速率生物降解为非毒性成分， （iii）生物催化物在设备的寿命中产生并在原位洗脱治疗剂。目标 该建议的是开发3D可打印树脂，这些树脂具有可调节的可生物降解水凝胶结构 刚度，并演示制造原型ELM设备以持续交付模型 化合物。在AIM 1中，我们将使用非致病性大肠杆菌Nissle 1917（ECN）创建水树脂，可以是 3D使用市售的3D打印机打印到水凝胶构造中。我们将制定水树脂 由可溶性球状蛋白衍生物组成，可以与水溶性丙烯酸酯共聚合 单体在光发射聚合时。 ELM水凝胶的机械性能（刚度， 每个树脂配方将量化强度，韧性）和酶促降解的速率。目标 2，我们将代谢设计非致病ECN，以产生berberine作为模型治疗化合物。 我们将进一步评估Caco-2细胞的细胞毒性和上皮完整性在3D印刷的情况下 榆树。作为对这些榆树的验证，我们将使用体外模型来确认Berberine的产生和洗脱 通过评估CACO-2对促炎刺激的反应，从3D打印的榆木中。我们设想这些 3D打印的榆树用作治疗恶性肿瘤或炎症的局部药物输送设备 影响肠道的疾病。虽然我们的ELM平台与广泛的微生物兼容 其中包括酵母和细菌，我们选择专注于大肠杆菌Nissle 1917，A的工程变体 肠道微生物组中常见的细菌的共生菌株。使用人类本地的微生物 微生物组可以促进我们平台的翻译以治疗肠道疾病。