We discuss our work in development of ‘Diffuse in vivo Flow Cytometry’ (DiFC) for non-invasive fluorescence enumeration of circulating tumor cells (CTCs) and describe recent progress towards human translation of DiFC. DiFC is an emerging technique wherein highly-scattered light is used to non-invasively sample blood flowing in large deep-seated blood vessels and detect fluorescently-labeled cells. The key advantages are that it allows continuous sampling of large circulating blood volumes and enumeration of rare cells over time. We discuss progress in development an application of near-infrared fluorescent molecular contrast agents for sensitive and specific labeling of CTCs directly in vivo. Candidate contrast agents include a folate receptor-targeted probe (OTL38, Cytalux), as well as new, purpose-designed pan-epithelial CTC-specific probes. We also discuss relevant tissue optics and instrumentation considerations for potential future human translation. Ultimately, DiFC could represent a new method for continuously enumerating CTCs without drawing blood samples that may enable early detection of cancer metastasis or monitoring of response to cancer therapies.
Cancer remains one of the leading causes of death worldwide despite advances in diagnostic and treatment approaches. Current methods of detection and diagnosis remain inaccessible or expensive in nature; therefore, the development of non-invasive strategies towards early-stage cancer detection are important to allow for early intervention, treatment, and access. Liquid biopsies have emerged as a non-invasive source to improve routine cancer monitoring, however cancer biomarker abundance is low, leading to limitations in detection and accuracy. The recent discovery of neoplastic circulating hybrid cells (CHCs) in peripheral patient blood provide the potential to improve detection sensitivity of blood-based assays using novel molecular-targeting contrast agents specific to both circulating tumor cells (CTCs) and CHCs. Additionally, these contrast agents can be detected using diffuse in vivo flow cytometry (DiFC), enabling non-invasive enumeration of cancer cell burden. Herein, the development and validation of near infrared (NIR) molecularly-targeted contrast agents with specificity towards epithelial biomarkers expressed on CHC and CTCs is discussed.
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