Modeling of human HSV infection: development of immune-competent 3D skin-on-chip with vascular perfusion

总结 传统的机理研究和药物开发严重依赖于体内动物模型和体外实验。 细胞培养系统尽管传统的体外培养的2D或3D细胞能够分析信号传导 途径和识别与感染或各种条件反应相关的标记基因 和治疗方法，它们不能重现组织中发生的复杂的细胞-细胞和细胞-基质相互作用， 微环境动物模型提供了对体内综合多器官反应的理解， 对它们的预测价值和对人类的生物学相关性存在严重关切。事实上，越来越多的 候选药物未能从I期临床试验进展到III期临床试验并进入市场。关键 减少临床试验中代价高昂的失败的挑战突出了产生更好的模型系统的紧迫性， 药物在人体中的有效性和安全性的临床前测试，以及了解分子机制 导致了成千上万的人类疾病一个三维（3D）人体组织模型承诺引人注目 预测复杂的生理功能，对传染病的免疫反应， 对治疗剂的药理学反应。这样的合成组织模型具有突出的潜力， 可靠的药物功效测试，并作为动物模型的上级替代品，特别是对于那些感染性 没有足够的动物模型。皮肤是人体最大的器官， 保护身体免受病原体和潜在有害物质渗透的屏障。为了 模拟人体器官水平的皮肤病理生理学，我们建议利用生物工程方法， 开发一种体外3D“芯片上的皮肤”，将循环免疫细胞纳入正常的 皮肤表皮和真皮用于模拟人单纯疱疹病毒（HSV）中病毒-宿主相互作用 感染我们的具体目标是：1）建立和验证血管化的3D“芯片上皮肤”平台， 供体来源的原代细胞用于HSV感染的建模和鉴定用于保护的关键免疫应答。 2)在3D血管化的“芯片上皮肤”中重述组织驻留记忆T细胞区室，用于建模 组织驻留记忆T细胞介导的局部免疫中的抗原特异性、细胞密度和TCR多样性 保护

SUMMARY Conventional mechanistic research and drug development relies heavily on in vivo animal models and in vitro cell culture systems. Although conventional in vitro cultured 2D or 3D cells enable the analysis of signaling pathways and the identification of signature genes associated with responses of infection or various conditions and treatments, they can't reproduce complex cell-cell and cell-matrix interactions occurring in the tissue microenvironment. Animal models provide understanding on in vivo integrated multi-organ responses but serious concerns exist over their predictive value and biological relevance to humans. In fact, more and more drug candidates have failed to advance from Phase I to Phase III clinical trials and to reach the market. Critical challenges to reduce costly failures in clinical trials highlight the urgency to generate better model systems for preclinical testing of drug efficacy and safety in humans, and for understanding molecular mechanisms underlying thousands of human diseases. A three-dimensional (3D) human tissue model promises compelling advantages to predict complex physiological functions, immune responses to infectious diseases, and pharmacological responses to therapeutic agents. Such a synthetic tissue model has outstanding potential for reliable drug efficacy testing and as a superior replacement for an animal model, especially for those infectious diseases that do not have adequate animal models. Skin is the largest organ of the human body and forms a barrier to protect the body against pathogens and penetration of potential harmful substances. In order to mimic the organ-level skin pathophysiology in humans, we propose to harness bioengineering approaches to develop an in vitro 3D `skin-on-chip' that incorporates circulating immune cells into the normal architecture of skin epidermis and dermis for modeling of viral-host interactions in human herpes simplex virus (HSV) infection. Our Specific Aims are: 1) Establish and validate a vascularized 3D `skin-on-chip' platform using donor-derived primary cells for modeling of HSV infection and identifying key immune responses for protection. 2) Recapitulate a tissue resident memory T-cell compartment in 3D vascularized `skin-on-chip' for modeling of antigenic specificity, cell density and TCR diversity in tissue resident memory T-cell mediated local immune protection.