Label-Free, Longitudinal, Multi-Metric Viability Imaging of 3D Tissue Spheroid Array

总结 当选择癌症治疗时，医生通常开始一线治疗方案，并监测患者的病情。 在观察和等待的基础上取得进展，遵循一套基于大型患者临床试验的指南 人口但这种传统的方法受到质疑，因为它是否为个别患者提供了 最佳治疗为了更好地找到一个匹配的治疗方案，一个叫做精准癌症医学的概念 或者个性化的癌症治疗已经被研究过了。在其他方法中，功能性精准医学 直接测试从病人身上活检的肿瘤细胞的化疗选择，以找到最佳匹配的治疗方法。 对于特定的病人。 然而，这种有前途的方法尚未被临床医生广泛采用，因为肿瘤 实验室中的微环境与患者体内的微环境不同，导致药物不一致。 由于样品和患者之间的反应，活检细胞的数量通常不足以进行检测， 可靠数量的选项进行测试。第一个问题正在解决的最新进展，在三个- 三维（3D）细胞培养技术，可以更好地模拟实验室中人体的微环境。但 第二个问题是可测试选项的数量有限，这主要是由于当前分析的局限性 在3D培养中评估化学敏感性的技术。使用大多数当前的检测方法， 一次，具有不同作用机制的多种药物不能同时由一个单一的测试。 比色法结合起来，这些限制指数地减少了可测试选项的数量， 多个评估时间点以设计序贯治疗或增加待测药物数量时 联合治疗 在这里，我们将开发一种新的技术，用于在3D培养中评估化疗敏感性，通过最大化 一种无标记3D显微技术的潜力，称为光学相干断层扫描（OCT）。大多数 先前的OCT研究仅测量了一种或两种类型的信号，并显示了对应于 在每项研究中只有单一类型的细胞活力破坏过程。但这种做法引发了一种担忧 关于特异性（即，除研究中测试的过程之外的其他类型的过程可以产生类似的OCT 信号）。这种低特异性，沿着不清楚的生存能力评估机制，阻碍了OCT 方法被采用的有前途的概念功能精准医学。 因此，我们将开发至少18种不同类型的OCT信号，并确定其灵敏度和 四种主要类型的生存能力破坏过程的特异性。这种方法的可行性已经得到 在一项初步研究中，我们对6,000多个3D培养的细胞进行了成像和分析， 球状体这个R 01项目将对多达100，000+个球体进行成像和分析， 研究OCT信号类型的全面范围。

SUMMARY When selecting cancer therapy, physicians generally begin with first-line treatment options and monitor patient progress on a watch-and-wait basis, following a set of guidelines based on clinical trials from a large patient population. But this traditional method has been questioned on whether it provides individual patients with the optimal treatment. To better find a matching treatment individually, a concept called precision cancer medicine or personalized cancer medicine has been studied. Among other approaches, functional precision medicine directly tests chemotherapy options on tumor cells biopsied from a patient to find the best matching treatment for the specific patient. This promising approach, however, has not been widely adopted by clinicians because the tumor microenvironment in a lab differed from the one within the patient’s body, leading to inconsistent drug responses between the sample and patient, and the quantity of biopsied cells is generally insufficient for a reliable number of options to be tested. The first problem is being addressed by recent advances in three- dimensional (3D) cell culture techniques, which better mimic the body’s microenvironment in a lab. But the second problem, the limited number of testable options, is mainly due to limitations in the current assay techniques that assess chemosensitivity in 3D culture. With most current assays, a sample can only be tested once, and multiple drugs with different mechanisms of action cannot be simultaneously tested by a single assay. Combined, these limitations exponentially reduce the number of testable options when involving multiple assessment time points to design a sequential therapy or when increasing the number of drugs to test a combination therapy. Here, we will develop a new technique for the assessment of chemosensitivity in 3D culture, by maximizing the potential of a label-free 3D microscopy technology, called optical coherence tomography (OCT). The majority of prior OCT research measured only one or two types of signals and showed the signals corresponding to only a single type of cell viability disruption process in each study. But this approach has led to a concern about specificity (i.e., other types of processes than the one tested in the study can generate similar OCT signals). This low specificity, along with unclear mechanisms of viability assessment, have prevented OCT methods from being adopted for the promising concept of functional precision medicine. Therefore, we will develop at least 18 different types of OCT signals and establish their sensitivity and specificity to four major types of viability disruption processes. The feasibility of this approach has been strongly supported by a pilot study where we imaged and analyzed more than 6,000 3D-cultured cell spheroids. This R01 project will image and analyze up to 100,000+ spheroids for an unprecedentedly systemic investigation of the comprehensive range of OCT signal types.