3D Bioprinted skin models for drug screening

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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 identified a 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. Jia 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. We have implemented an HTS of a library of 700 compounds. A selected number of active compounds from the HTS are now being 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 have now completed screens of an oncology collection of 900 compounds to identify new chemotherapeutic agents for these cancers in a 3D tissue model. We have initiated a project to incorporate human macrophages into biofabricated skin tissue models to investigate the effect of immune cells in skin diseases, including fibrosis, wound healing and others. 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. 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. We have initiated a collaboration with Dr. Kyung Sung at the FDA/CBER to assess the therapeutic effects of mesenchymal stem cells in a biofabricated skin model of atopic dermatitis.