Fluorescence molecular imaging using ABY-029, an epidermal growth factor receptor (EGFR)-targeted synthetic Affibody peptide labeled with a near-infrared fluorophore, is under investigation for surgical guidance during head and neck squamous cell carcinoma (HNSCC) resection. However, tumor-to-normal tissue contrast is confounded by intrinsic physiological limitations of heterogeneous EGFR expression. In this study, a machine learning-based optomics analysis, which interprets the textural pattern differences in EGFR expression conveyed by fluorescence, was applied to optical ABY-029 fluorescence image data of HNSCC surgical specimens. The study objective was to determine the correlations between optomics method classification performance and tissue inherent EGFR expression level. Fluorescence image data were collected through a Phase 0 clinical trial of ABY-029, which involved a total of 20,073 sub-image patches (size of 1.8×1.8 mm2) extracted from 24 bread-loafed slices of HNSCC surgical resections from 12 patients who were stratified into three dose groups (30, 90, and 171 nanomoles). The optomics approach utilized a supervised machine learning pipeline. Each dose group was randomly partitioned on the specimen-level 75%/25% into training/testing sets, then all training and testing sets were aggregated. A total of 1,472 standardized optomic features were extracted from each patch and evaluated by minimum redundancy maximum relevance feature selection, and 25 top-ranked features were used to train a support vector machine classifier. A conceptual framework of correlation analysis to evaluate the relationship between optomics tumor classification performance and underlying EGFR expression level was provided, but the present results are underpowered. Some generalized conclusions about the ABY-029 fluorescence optomics method correlating to varied levels of EGFR expression were summarized, suggesting that optomics method using fluorescence molecular imaging data offers a potentially stable image analysis technique for cancer detection for fluorescence-guided surgery applications; however, further study with additional samples is needed to validate this conclusion.
During resection of head and neck cancers, surgeons collect multiple small tissue samples from the excision wound bed for “frozen section” analysis by a clinical pathologist to determine clear margins. However, this process is disruptive to work flow, time-consuming, and expensive. We have validated Paired-Agent Imaging (PAI) in in vivo xenograft models using epidermal growth factor receptor and demonstrated that PAI exhibits higher diagnostic accuracy targeted agent alone – tested in both fresh tissue (~3-mm) and frozen sections (10-m). Here PAI will be extended to screening of tissue biopsies to quantify tumor burden in comparison to standard frozen section pathology.
Short-wave infrared (SWIR/NIR-II) fluorescence imaging has received increased attention for use in fluorescence-guided surgery (FGS) due to the potential for higher resolution imaging of subsurface structures and reduced autofluorescence compared to conventional NIR-I imaging. As with any fluorescence imaging modality introduced in the operating room, an appropriate accounting of contaminating background signal from other light sources in the operating room is an important step. Herein, we report the background signals in the SWIR and NIR-I emitted from commonly-used equipment in the OR, such as ambient and operating lights, LCD screens and surgical guidance systems. These results can guide implementation of protocols to reduce background signal.
ABY-029, an anti-epidermal growth factor receptor (EGFR) Affibody molecule labeled with IRDye 800CW, has been used in three Phase 0 microdosing clinical trials for fluorescence guided surgery. In May of 2019, the clinical trials were put on hold because the ABY-029 produced under Good Manufacturing Practices (GMP) for human administration had come to the end of term in which the drug product was known to be stable. Stability testing was halted due to limitations in supply of a suitable reference standard and a required test product being discontinued from commercial sale. In order to complete the remaining patients in the three clinical trials, new stability tests were developed and the GMP batch of ABY- 029 drug product tested under the new protocols. The GMP batch of ABY-029 passed all stability tests under the new protocols and the Federal Drug Administration (FDA) has given permission to complete the remaining patients with stability testing of ABY-029 performed for each patient. The tests developed and used to test ABY-029 drug product stability are described here.
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