3D Bioprinted skin models for drug screening

为响应RFA-TR-19-020：使用生物浓缩的三维皮肤病组织模型(U18)进行药物筛选，正在实施两个项目： 1)与哥伦比亚大学的安吉拉·克里斯蒂亚诺博士合作，利用3D生物打印技术将人类原始组织作为一种疾病模型进行药物发现。我们已经成功地开发了96孔跨孔板形式的全厚皮肤等价物(真皮和表皮)的生物制造协议，这将使化合物筛选产量显著增加。我们一直在探索不同的细胞因子鸡尾酒，它们模拟牛皮癣的压力，并在皮肤组织上产生牛皮癣的表型。细胞因子分泌试验将被用作牛皮癣的表型分析，并用于目前计划的筛查。 2)与华盛顿大学朱继智博士合作建立了用于抗人类单纯疱疹病毒感染的抗病毒药物的生物浓缩3-D皮肤模型。一个高通量筛选，以确定潜在的抗病毒化合物，以阻止HSV感染在生物丰富的3-D全厚皮肤模型。我们使用HSV-GFP报告病毒感染96孔板形式的生物打印的全厚皮肤等价物，并使用荧光显微镜测量病毒感染和复制。一个有700个化合物的库的高温超导已经实现。候选的抗病毒药物将在血管3D皮肤模型中得到验证，该模型由患者特有的原始角质形成细胞、成纤维细胞和内皮细胞构建，这些细胞来自不同的HSV结果队列。 我们继续使用生物打印的全厚皮肤等效物和荧光标记的癌细胞，以96孔板的形式建立皮肤癌模型(皮肤鳞状细胞癌和黑色素瘤)。分析表明，HTS的检测是可靠的，我们现在正在扩大筛选集中的药物集合，以确定这些癌症的新化疗药物。 我们正在与NIAID的Yasmine Belkaid博士的实验室合作，通过测量炎症细胞因子的分泌来探索微生物群对血管化皮肤等价物上的皮肤免疫反应的影响。 最后，我们与NCATS的Tox21小组建立了合作关系，由FDA/NCTR的夏梦航博士和Luisa Camacho博士领导，探索使用生物软化的皮肤组织来研究药物的渗透性。

In response to the RFA-TR-19-020: Drug Screening with Biofabricated 3-D Skin Disease Tissue Models (U18), two projects are being implemented: 1) 3D Bioprinting of human native-like tissues as disease-in-a-dish models for drug discovery with Dr. Angela Christiano at Columbia University. We have been successfully developed biofabrication protocols for full thickness skin equivalents (with dermis and epidermis) in a 96-well transwell plate format which will enable a significance increase in compound screening throughput. We have been exploring different cocktails of cytokines that mimic a psoriatic stress and produces a psoriatic phenotype on the skin tissue. A cytokine secretion assay will be used as a phenotypic assay for psoriasis and used for the screen which is now planned. 2) Biofabricated 3-D skin model for antiviral drug discovery against human HSV infection with Dr. Jiz Zhu at University of Washington. A High-throughput screen to identify potential antiviral compounds that block HSV infection in biofabricated 3-D full thickness skin models. We have used a HSV-GFP reporter virus to infect a bioprinted full thickness skin equivalents in a 96-well plate format and measured viral infection and replication using fluorescence microscopy. The HTS of a library of 700 compounds have been implemented. Candidate antivirals will be validated in a vascularized 3-D skin models fabricated with patient-specific primary keratinocyte, fibroblast and endothelium cells from a cohort of diverse HSV outcomes. We have continued to develop skin cancer models (cutaneous squamous cell carcinoma and melanoma) using bioprinted full thickness skin equivalents and fluorescently labeled cancer cells, in a 96-well plate format. The assays have shown to be robust for HTS and we are now scaling up for screening of a focused collection of drugs to identify new chemotherapeutic agents for these cancers. We have an on-going collaboration with the laboratory of Dr. Yasmine Belkaid at NIAID exploring the effects of microbiome on skin immune responses on vascularized skin equivalents by measuring secretion of inflammatory cytokines. Finally, we have established a collaboration with the Tox21 group at NCATS led by Dr. Menghang Xia and Dr. Luisa Camacho at the FDA/NCTR, to explore the use of biofabricated skin tissues to study drug permeability.