3D Bioprinting of human native-like tissues as disease-in-a-dish models for drug discovery

人类天然组织的 3D 生物打印作为药物发现的皿中疾病模型

• 批准号: 9355001
• 负责人: Marc Ferrer
• 金额: $ 301.07万
• 依托单位: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/9355001
• 关键词: 3D Print; Adipocytes; Affect; Age related macular degeneration; Aging; Animal Model; Architecture; Back; Biological Assay; Blindness; Blood Vessels; Blood-Retinal Barrier; Body part; Cancer Cell Growth; Cancer Patient; Cardiovascular Diseases; Cardiovascular system; Cell Differentiation process; Cell physiology; Cells; Cellular biology; Cessation of life; Chemical Agents; Chicago; Child; Clinical; Clinical Trials; Collaborations; Complex; Coronary Arteriosclerosis; Culture Techniques; Degenerative Disorder; Dermal; Dermis; Developed Countries; Development; Developmental Biology; Disease; Disease Progression; Disease model; Electrophysiology (science); Environment; Epidermis; Etiology; Event; Eye; Failure; Fibroblasts; Functional disorder; Future; Genes; Genetic; Goals; Hereditary Disease; Histocompatibility Testing; Histology; Human; Hydrogels; Immune; In Vitro; Individual; Leukocytes; Libraries; Life Expectancy; Link; Malignant Neoplasms; Malignant neoplasm of ovary; Maryland; Medial; Methodology; Methods; Microscopy; Modeling; Molecular; Morphology; National Human Genome Research Institute; Neoplasm Metastasis; Omentum; Operative Surgical Procedures; Pathology; Patients; Pattern; Pharmaceutical Preparations; Pharmacological Treatment; Pharmacology; Phenotype; Photoreceptors; Physiological; Physiology; Plug-in; Preclinical Drug Evaluation; Predictive Value; Printing; Process; Progeria; Proteins; Protocols documentation; Research Personnel; Resources; Retina; Retinal Degeneration; Site; Skin; Skin Tissue; Smooth Muscle Myocytes; Software Tools; Staging; Stroke; Structure; Structure of retinal pigment epithelium; Syndrome; System; Techniques; Technology; Therapeutic; Tissue Engineering; Tissue Model; Tissues; Toxic effect; United States National Institutes of Health; Universities; Vascular Diseases; Vision; Water; age related; biocompatible polymer; bioprinting; cell type; college; data exchange; drug discovery; human tissue; in vivo; induced pluripotent stem cell; insight; keratinocyte; neoplastic cell; prevent; programs; screening; senescence; skin disorder; stem cell technology; stressor; therapeutic development; three-dimensional modeling; tissue biomarkers; tumor; tumor microenvironment

The activities of the Tissue Bioprinting program include: i) Develop bioprinting protocols to fabricate native-like, functional human tissues. Ii) Validate of printed tissues: develop morphological and physiological biomarkers of tissue architecture and function by using microscopy, histology, gene sequencing, electrophysiological and other methodologies. iii) Use printed tissues for the screening of focused libraries of compounds for drug discovery. iv) Develop a framework for the sharing of validated protocols as a readily available resource for researchers to exchange data on optimized conditions for bioinks, culture techniques, cell types, and software tools as well as techniques to quantify and validate printed tissues. Bioprinting of dermal tissue for modeling skin diseases: Skin is a complex, hierarchical and stratified tissue that provides protection from the external environment by acting as an active physical barrier into the body and regulating transport of water and other metabolites out of the body. Bioprinted human native skin models can provide fundamental insights into the etiology of skin diseases as well as elucidate the pathophysiological mechanisms in skin disease progression and discovery of treatments. Current efforts are directed to the bioprinting of consistent and reproducible human native-like dermis and epidermis layers of the skin using normal human fibroblasts and keratinocytes. Once this is achieved, we will introduce available disease patient cells to reproduce disease skin tissues that will be used as disease-in-dish models for screening. Bioprinting of blood retinal barrier for a disease model of wet age related macular degeneration (wet AMD): Retinal degenerative diseases are the leading cause of irreversible vision loss in developed countries. In many cases the diseases originate in the homeostatic unit in the back of the eye that contains the retina, retinal pigment epithelium (RPE) and the choriocapillaris. In diseases like age-related macular degeneration (AMD), it is thought that RPE dysfunctions cause disease-initiating events and as the RPE degenerates photoreceptors begin to die and patients start losing vision. Patient-specific induced pluripotent stem (iPS) cell-derived RPE provides direct access to a patients genetics and allow the possibility of identifying the initiating events of RPE-associated degenerative diseases. Three-dimensional models of the RPE, neuroretina, and the choriocapillaris are being developed using tissue bioprinting combined with iPS cell technology and fundamental developmental biology. Analysis of disease processes at the level of this entire homeostatic unit will likely provide more insight into molecular mechanisms of retinal degenerative diseases, as well as providing a native disease model for the discovery of new treatments for AMD. This is a collaboration with the group of Dr. Kapil Bharti at NEI. Bioprinting of a blood vessel wall model for modeling Progeria: Hutchinson-Gilford Progeria syndrome (HGPS) is a genetic disorder that, although rare, has devastating consequences to the affected children. Those with HGPS undergo accelerated aging and have an average life expectancy of just 13.4 years. Patients with HGPS suffer from accelerated vascular disease, and death almost always results from coronary artery disease or stroke. Previous studies have shown a massive loss of smooth muscle cells (SMCs) in the medial layer of large vessels in HGPS patients and animal models, suggesting a possible link between this SMC loss phenotype and the deadly cardiovascular malfunction associated with HGPS. 3D bioprinting techniques are being used to build and characterize a tissue engineered blood vessel wall system using HGPS and normal control iPSC-derived SMCs as disease-in-a-dish models for the discovery of treatments for HGPS. The 3D bioprinted tissue models of blood vessel walls in multi-well plates will be validated biologically and pharmacologically using treatment options previously described. We expect that this 3D vessel system of HGPS will be of great importance for future drug screening and therapeutic development for HGPS and age-related cardiovascular diseases, especially since the toxic protein that causes HGPS is also made in small quantities in normal individuals, and increases as cells approach senescence. This is a collaboration with Dr. Can Kao at University of Maryland, College Park, and Dr. Francis Collins, at NIH/NHGRI. Bioprinting of an omentum model for modeling ovarian cancer metastasis: Metastasis is the process of spreading of tumor cells to different parts of the body and in most cases it is the pathology that leads to ultimate death in cancer. A 3D assay that recreated the human omentum using cells from ovarian cancer patients undergoing surgery was successfully used to discover compounds that would prevent attachment of tumor cells to the omentum, an early site of metastasis in ovarian cancer. We are currently using tissue bioprinting techniques to increase the relevance on the ovarian cancer metastasis omentum model by introducing additional cell types that are important for the tumor-microenvironment interaction in the omentum metastatic site, including vasculature, adipose cells and leukocytes. Once a native omentum model is recreated, we will study the growth of cancer cells and screen for pharmacological agents that prevent tumor metastasis. This is a collaboration with Dr. Ernst Lengyels group at the Universtiy of Chicago.

组织生物打印计划的活动包括：i)开发生物打印方案，以制造天然的、有功能的人体组织。Ii)印刷组织的验证：利用显微镜、组织学、基因测序、电生理学等方法，开发组织结构和功能的形态和生理生物标记物。三)使用印刷组织筛选用于药物发现的重点化合物文库。四)开发一个共享有效方案的框架，作为研究人员随时可用的资源，以交流关于生物墨水、培养技术、细胞类型、软件工具以及量化和验证打印组织的技术的优化条件的数据。 用于模拟皮肤病的真皮组织生物打印：皮肤是一种复杂的、分层的组织，通过充当进入身体的主动物理屏障并调节水分和其他代谢物的运输来提供对外部环境的保护。生物打印的人类天然皮肤模型可以为皮肤病的病因学提供基本的见解，并阐明皮肤病进展和发现治疗方法的病理生理机制。目前的努力是使用正常的人成纤维细胞和角质形成细胞来打印一致的和可重复的人类自然样真皮和表皮层的生物打印。一旦实现了这一点，我们将引入可用的疾病患者细胞来复制疾病皮肤组织，这些组织将被用作筛查疾病的模型。 湿性老年性黄斑变性(湿性AMD)疾病模型的血视网膜屏障生物打印：在发达国家，视网膜退行性疾病是不可逆性视力丧失的主要原因。在许多情况下，疾病起源于眼后部的动态平衡单位，该单位包含视网膜、视网膜色素上皮(RPE)和脉络膜毛细血管。在像老年性黄斑变性(AMD)这样的疾病中，人们认为RPE功能障碍会导致疾病的发生，随着RPE的退化，光感受器开始死亡，患者开始失明。患者特异性诱导多能干细胞(IPS)来源的RPE提供了对患者遗传学的直接访问，并使识别RPE相关退行性疾病的启动事件成为可能。利用组织生物打印技术结合iPS细胞技术和基础发育生物学，正在开发RPE、神经视网膜和脉络膜毛细血管的三维模型。在整个动态平衡的水平上对疾病过程进行分析，可能会为视网膜退行性疾病的分子机制提供更多的见解，并为发现AMD的新治疗方法提供一个天然的疾病模型。这是与NEI的Kapil Bharti博士团队的合作。 用于模拟早衰症的血管壁模型的生物打印：Hutchinson-Gilford Progeria综合征(HGPS)是一种遗传性疾病，虽然罕见，但对受影响的儿童具有毁灭性的后果。患有HGP的人会加速衰老，平均预期寿命只有13.4岁。HGP患者患有加速的血管疾病，死亡几乎总是由冠状动脉疾病或中风引起。以前的研究表明，HGPS患者和动物模型大血管中层的平滑肌细胞(SMC)大量丢失，提示这种SMC丢失表型与HGPS相关的致命心血管功能障碍之间可能存在联系。3D生物打印技术正被用于构建和表征组织工程血管壁系统，该系统使用HGPS和正常对照IPSC来源的SMC作为发现HGPS治疗方法的盘中病模型。多孔板中血管壁的3D生物打印组织模型将使用前面描述的治疗方案进行生物学和药理学验证。我们预计，这种HGPS的3D血管系统将对未来HGPS和年龄相关心血管疾病的药物筛选和治疗开发具有重要意义，特别是因为导致HGPS的有毒蛋白在正常人中也有少量产生，并随着细胞接近衰老而增加。这是与马里兰大学学院公园的Can Kao博士和NIH/NHGRI的Francis Collins博士的合作。 模拟卵巢癌转移的大网膜模型的生物打印：转移是肿瘤细胞扩散到身体不同部位的过程，在大多数情况下，它是导致癌症最终死亡的病理因素。一项使用卵巢癌手术患者细胞重建人大网膜的3D测试成功地被用来发现可以防止肿瘤细胞附着到大网膜的化合物，大网膜是卵巢癌早期转移的部位。我们目前正在使用组织生物打印技术，通过引入其他类型的细胞，包括血管系统、脂肪细胞和白细胞，来增加与卵巢癌转移大网膜模型的相关性，这些细胞类型对于大网膜转移部位的肿瘤-微环境相互作用至关重要。一旦重建了天然的大网膜模型，我们将研究癌细胞的生长情况，并筛选防止肿瘤转移的药物。这是与芝加哥大学Ernst Lengyels博士团队的合作。