Disease-homing light delivery by engineering bioluminescent immune cells for whole body precision photomedicine

项目总结 光动力疗法(PDT)提供了独特的细胞死亡机制，并被用于治疗多种癌症。 诊所。光疗的原理是将光活性化学物质运送到病区转化为事故 将光子能量转化为局部化学毒性，避免全身电击。PDT对经典药物是不可知的- 耐药途径，不会导致严重的共病，使其作为一种单一疗法或作为一种 多模式治疗的组成部分。然而，对PDT的主要批评是需要外部光源， 这将PDT的应用限制在表面病变或通过光纤可到达的病变。为了克服这一点， PDT的光传递机制最近出现了一种范式转变：生物发光 介导性光动力疗法(BL-PDT)，其中BL酶激活光谱匹配的、共定位的光敏剂(PS) 在病变内，消除了对外部光源的需要。尽管这克服了一个主要障碍， 目前利用半导体纳米结构传递BL酶的方法在能力上是有限的 对这种疾病进行局部定位。在这里，我们提出了一种新的BL酶传递和BL-PDT的方法：TO 基因工程超亮生物发光免疫细胞(UBLI)及其疾病归宿 像趋化性一样，在疾病部位进行交通和积累的能力。这种方法不再需要 对于作为药物输送组件的复杂和潜在有毒的纳米结构-只有BL底物和 将需要PS给药(两种无毒化合物)。最近，我们引入了精度 使用靶向、可激活的PS进行光医学治疗，表现出细胞选择性并减少靶外毒性 在转移性癌症模型中。我们将把拟议的新型光传输平台与精确度配对 为最大利益而进行的光医学。首先，我们将优化BL-PDT平台在3D肿瘤-免疫细胞联合 由蒙特卡罗模拟提供信息的生物和临床相关参数空间的培养。 然后，我们将通过静脉注射UBLIS到体内异种移植体内来演示该方法 体外实验结果所提供的肿瘤转移模型。这些概念验证研究将使 在未来的资助期，在多种疾病模型中进行全面的安全性和有效性研究。最终，我们 设想临床翻译涉及患者免疫细胞的提取和工程，类似于嵌合体 抗原重定向(CAR)T细胞治疗，然后回输和给药。这是一项新的 治疗模式有可能惠及癌症及其他领域的许多疾病，这证明了高- 这项提议的风险、高回报性质。

PROJECT SUMMARY Photodynamic therapy (PDT) offers unique mechanisms of cell death and is used to treat many cancers in the clinic. The principal of phototherapy is that photoactive chemicals delivered to the disease site convert incident photonic energy into local chemical toxicity, avoiding a systemic shock. PDT is agnostic to classical drug- resistance pathways and does not cause critical co-morbidities, making it attractive as a monotherapy or as a component of multi-modal therapy. However, a major criticism of PDT is the need for an external light source, which limits the application of PDT to superficial lesions or those accessible by fiber optics. To overcome this, a paradigm shift in the mechanism of light delivery for PDT has recently emerged: bioluminescence (BL) mediated PDT (BL-PDT), wherein BL enzymes activate spectrally matched, co-localized photosensitizers (PS) within the lesion, eliminating the need for an external light source. Although this overcomes a major hurdle, current methods that leverage semiconductor nanoconstructs for BL enzyme delivery are limited in their ability to localize to the disease. Here, we propose a novel method for BL enzyme delivery and BL-PDT: to genetically engineer ultra-bright bioluminescent immune cells (UBLIs) and exploit their disease-homing capabilities, like chemotaxis, to traffic to and accumulate in sites of disease. This approach eliminates the need for complex and potentially toxic nanoconstructs as a component of drug delivery—only the BL substrate and PS administration (both non-toxic compounds) will be required. Recently, we introduced precision photomedicine using a targeted, activatable PS that exhibited cellular selectivity and reduced off-target toxicity in a metastatic cancer model. We will pair the proposed novel light delivery platform with precision photomedicine for maximal benefit. First, we will optimize the BL-PDT platform in 3D cancer–immune cell co- cultures across biologically and clinically relevant parameter spaces informed by Monte Carlo simulations. Then we will demonstrate the approach in vivo by intravenous injection of UBLIs into an in vivo xenograft model of cancer metastases informed by in vitro results. These proof-of-concept studies will enable comprehensive safety and efficacy studies in multiple disease models in future funding periods. Ultimately, we envision clinical translation involving extraction and engineering of patient immune cells, similar to chimeric antigen redirected (CAR) T cell therapy, followed by reinfusion and administration of photomedicine. This new therapeutic paradigm has potential to benefit many diseases in cancer and beyond, which justifies the high- risk, high-reward nature of the proposal.