Fluorescein videoangiography data analysis protocol for mapping retinal vascular permeability in humans.
Fluorescein videoangiography data analysis protocol for mapping retinal vascular permeability in humans.
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用于绘制人类视网膜血管通透性的荧光素视频血管造影数据分析协议。
DOI：
10.1117/12.2650304
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Tichauer,KennethM
中科院分区：
文献类型：
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作者：
Vavrek,Sarah;Kayaalp-Nalbant,Elif;Konopek,Nicholas;Bou-Ghanem,Ghazi;Fawzi,AmaniA;Mieler,WilliamF;Kang-Mieler,JenniferJ;Tichauer,KennethM
结合动力学在癌症诊断和治疗中起着重要作用。然而，目前的定量结合动力学的方法未能考虑药物和成像剂在生物组织中经历的三维环境。作为回应，使用配对试剂分子成像原理开发了一种在3-D组织培养中测定试剂结合和解离的方法。为了测试该方法，在整个染色和冲洗过程中，在四种不同的人癌细胞系中测量ABY-029（IRDye 800 CW标记的表皮生长因子受体（EGFR）靶向抗体模拟物）和IRDye-700DX羧酸盐在3-D球状体中的摄取。然后将隔室模型（针对应用优化）拟合到两种成像剂的动力学曲线以估计EGFR靶向ABY-029剂的结合和解离速率常数。在实验和模拟中观察到表观结合速率常数（k3）与受体浓度之间的统计学显著相关性（r= 0.99，p < 0.05）。对于具有不同EGFR表达水平的细胞系，在有效k 3（ABY-029结合EGFR的表观速率常数）值之间发现统计学显著差异（p< 0.05），对于其他拟合参数，在细胞系和对照之间没有发现显著差异。此外，与金标准方法相比，通过该模型确定了相似的结合亲和力曲线。这种在临床相关的3-D肿瘤球体模型中量化成像剂或药物结合亲和力的低成本方法可用于指导分子引导手术中的成像时机，并可能对药物开发产生影响。
Binding kinetics play an important role in cancer diagnosis and therapeutics. However, current methods of quantifying binding kinetics fail to consider the three-dimensional environment that drugs and imaging agents experience in biological tissue. In response, a methodology to assay agent binding and dissociation in 3-D tissue culture was developed using paired-agent molecular imaging principles. To test the methodology, the uptakes of ABY-029 (an IRDye 800CW-labeled epidermal growth factor receptor (EGFR)-targeted antibody mimetic) and IRDye-700DX carboxylate in 3-D spheroids were measured in four different human cancer cell lines throughout staining and rinsing. A compartment model (optimized for the application) was then fit to the kinetic curves of both imaging agents to estimate binding and dissociation rate constants of the EGFR-targeted ABY-029 agent. A statistically significant correlation was observed between apparent association rate constant (k3) and the receptor concentration experimentally and in simulations (r= 0.99,p< 0.05). A statistically significant difference was found between effectivek3(apparent rate constant of ABY-029 binding to EGFR) values for cell lines with varying levels of EGFR expression (p< 0.05), with no significant difference found between cell lines and controls for other fit parameters. Additionally, a similar binding affinity profile compared to a gold standard method was determined by this model. This low-cost methodology to quantify imaging agent or drug binding affinity in clinically relevant 3-D tumor spheroid models can be used to guide timing of imaging in molecular guided surgery and could have implications in drug development.