3D Bioprinted skin models for drug screening

为了响应 RFA-TR-19-020：使用生物制造的 3-D 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 Angela Christiano 博士一起对人类天然组织进行 3D 生物打印，作为药物发现的皿中疾病模型。 我们已经成功开发了 96 孔 Transwell 板格式的全层皮肤等效物（包含真皮和表皮）的生物制造方案，这将显着提高化合物筛选通量。 我们一直在探索模拟银屑病应激并在皮肤组织上产生银屑病表型的不同细胞因子混合物。 细胞因子分泌测定将用作银屑病的表型测定，并用于目前计划的筛选。 2) 与华盛顿大学的 Jiz Zhu 博士合作开发生物制造的 3D 皮肤模型，用于发现人类 HSV 感染的抗病毒药物。高通量筛选，用于识别生物制造的 3D 全厚皮肤模型中阻断 HSV 感染的潜在抗病毒化合物。 我们使用 HSV-GFP 报告病毒感染 96 孔板形式的生物打印全层皮肤等效物，并使用荧光显微镜测量病毒感染和复制。 700 种化合物的库的 HTS 已经实施。候选抗病毒药物将在血管化 3D 皮肤模型中进行验证，该模型由来自一组不同 HSV 结果的患者特异性原代角质形成细胞、成纤维细胞和内皮细胞制成。 我们继续使用生物打印的全层皮肤等效物和荧光标记的癌细胞以 96 孔板形式开发皮肤癌模型（皮肤鳞状细胞癌和黑色素瘤）。 这些检测方法对于 HTS 来说是稳健的，我们现在正在扩大筛选重点药物集合，以确定针对这些癌症的新化疗药物。 我们与 NIAID 的 Yasmine Belkaid 博士实验室持续合作，通过测量炎症细胞因子的分泌，探索微生物组对血管化皮肤等同物的皮肤免疫反应的影响。 最后，我们与FDA/NCTR的Menghang Xia博士和Luisa Camacho博士领导的NCATS Tox21小组建立了合作，探索使用生物制造的皮肤组织来研究药物渗透性。

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.