3D Bioprinted skin models for drug screening

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In response to the RFA-TR-17-007: NCATS Pilot Program for Collaborative Drug Discovery Research using Bioprinted Skin Tissue (U18), two projects were on boarded: 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. In addition, 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) Non-destructive, high throughput cytometry for drug discovery using trimodal confocal images of novel, 3D printed skin carcinoma construct with Drs.Dan Gareau at Rockefeller University and John Carucci at NYU. We have developed a skin squamous cell carcinoma (SCC) by introducing fluorescence labeled SCC tumor cells in the full thickness skin model and monitoring tumor growth using epifluorescence. Histological, microscopic and transcriptomics analysis showed that the SCC skin tissue model mimic many features of the native tumors. In addition, treatment of the SCC skin tissue with 5-fluorouracil (5-FU), standard of care for skin SCC, demonstrated efficacy in reducing tumor number and size in the tissue. We also collaborated with Dr. Ian Myles at NIAID to demonstrate the use of biofabricated skin tissues to demonstrate corrective effects Roseomonas mucosa treatments of atopic dermatatis phentypes. This contribution was part of a larger body of work by Dr. Myles lab that demonstarted that therapeutic responses to Roseomonas mucosa in atopic dermatitis involve lipid-mediated TNF-related epithelial repair. We have an on-going collaboration with the laboratory of Dr. Yasmine Belkaid at NIAID exploring the effects of microbiome on skin immune responses. Finally, we established a collaboration with the Tox21 group at NCATS led by Dr. Menghang Xia to explore the use of biofabricated skin tissues and a multiplex readout scheme (cell viability, TEER and cytokine secretion) to assess the irritation potential of compounds used in skin products. Forty-six toxic compounds identified from an initial screen with the monolayer culture systems were further tested for skin irritation potential on reconstructed human epidermis (RhE) and full thickness skin (FTS) three-dimensional (3D) tissue model constructs. Skin irritation potential of the compounds was assessed by measuring tissue viability, trans-epithelial electrical resistance (TEER), and secretion of cytokines interleukin 1 alpha (IL-1) and interleukin 18 (IL-18). Among known irritants, high concentrations of methyl violet and methylrosaniline decreased viability, lowered TEER, and increased IL-1 secretion in both RhE and FTS models, consistent with irritant properties. However, at low concentrations, these two compounds increased IL-18 secretion without affecting levels of secreted IL-1, and did not reduce tissue viability and TEER, in either RhE or FTS models. This result suggests that at low concentrations, methyl violet and methylrosaniline have an allergic potential without causing irritation. Using both HTS-compatible 2D cellular and 3D tissue skin models, together with irritation relevant activity endpoints, we obtained data to help assess the irritation effects of topical-use compounds and identify potential dermal hazards.