Bioprinting 3D skin for patient-specific drug discovery in inflammatory skin diseases.

项目摘要 银屑病是一种炎症性皮肤病，伴有异常的表皮过度增殖，影响2%-3%的 美国人口。目前还没有可靠的体外模型来证明银屑病药物的有效性和安全性。 考生可以接受测试。为了满足这一重大的未得到满足的药品需求，我们将结合IPSC 重新编程和3D生物打印技术构建3D皮肤模型以筛选治疗此病的药物 疾病。将使用IPSC方法，因为IPSC具有无限的增殖潜力，并且 能够分化成不同的细胞系，这使我们能够绕过 可以从患者那里获得的原代细胞的可用性有限。我们成功地构建了3D皮肤 仅使用IPSC来源的细胞(成纤维细胞和角质形成细胞)，证明了这一技术的可行性 接近。3D生物打印可以让我们重复构建复杂的结构来重现人类 组织/器官。在这个项目中，我们将首先用原代角质形成细胞构建具有免疫功能的3D皮肤， 成纤维细胞和T细胞。然后，IPSCs将从成纤维细胞或从正常组织中获得的血细胞中重新编程 和牛皮癣患者。角质形成细胞和成纤维细胞将从IPSCs分化为3D 皮肤模型，而T细胞的大量可获得性使我们能够直接使用主要来源的细胞。 生物印染的牛皮癣特异性IPSC来源的皮肤结构将用于疾病建模和 药理验证。CRISPR技术将用于将荧光记者集成到IPSC衍生的 角质形成细胞用于监测银屑病的发病诱因和药物治疗效果。本项目竣工 将为我们提供一个可靠的3D生物打印模型来开发治疗牛皮癣的药物。此外，我们的3D皮肤 模型可以很容易地调整和修改，以筛选治疗其他皮肤病的药物，使用一个平台 接近。

Project Summary Psoriasis is an inflammatory skin disorder with abnormal epidermal hyperproliferation, which affects 2-3 % of the US population. There are no reliable in vitro models with which the efficacy and safety of psoriasis drug candidates can be tested. To meet this major unmet pharmaceutical need, we will combine iPSC reprograming and 3D bioprinting technologies to construct a 3D skin model to screen for drugs to treat this disease. iPSC methodology will be used, because iPSCs have unlimited proliferation potential and the capability to differentiate into different cell lineages, which allows us to circumvent the limitation imposed by the limited availability of primary cells that can be procured from patients. We successfully constructed 3D skin using exclusively iPSC-derived cells (fibroblasts and keratinocytes), demonstrating the feasibility of this approach. 3D bioprinting can allow us to reproducibly construct complex structures to recapitulate human tissues/organs. In this project, we will first construct immunocompetent 3D skin with primary keratinocytes, fibroblasts and T cells. Then, iPSCs will be reprogramed from fibroblasts or blood cells obtained from normal and psoriatic individuals. Keratinocytes and fibroblasts will be differentiated from iPSCs for construction of 3D skin models, whereas the abundant availability of T cells allows us to use cells directly from primary sources. Bioprinted psoriasis-specific iPSC derived skin constructs will be used for disease modeling and pharmacological validation. CRISPR technology will be used to integrate fluorescent reporters in iPSC-derived keratinocytes to monitor disease induction of psoriasis and effect of drug treatment. Completion of this project will provide us with a reliable 3D bioprinted model to develop drugs to treat psoriasis. Moreover, our 3D skin model can be easily adapted and modified to screen for drugs to treat other skin diseases, using a platform approach.