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Published on: September 19, 2014
Evaluation of a Centyrin-Based Near-Infrared Probe for Fluorescence-Guided Surgery of Epidermal Growth Factor
Sakkarapalayam M Mahalingam, Vadim Y Dudkin1, Shalom Goldberg1
1Janssen Research & Development , 1400 McKean Road, Springhouse, Pennsylvania 19477, United States.
Abstract:
Tumor-targeted near-infrared fluorescent dyes have the potential to improve cancer surgery by enabling surgeons to locate and resect more malignant lesions where good visualization tools are required to ensure complete removal of malignant tissue. Although the tumor-targeted fluorescent dyes used in humans to date have been either small organic molecules or high molecular weight antibodies, low molecular weight protein scaffolds have attracted significant attention because they penetrate solid tumors almost as efficiently as small molecules, but can be infinitely mutated to bind almost any antigen. Here we describe the use of a 10 kDa protein scaffold, a Centyrin, to target a near-infrared fluorescent dye to tumors that overexpress the epidermal growth factor receptor (EGFR) for fluorescence-guided surgery (FGS). We have developed and optimized the dose and time required for imaging small tumor burdens with minimal background fluorescence in real-time fluorescence-guided surgery of EGFR-expressing tumor xenografts in murine models. We demonstrate that the Centyrin-near-infrared dye conjugate (CNDC) binds selectively to human EGFR+ cancer cells with an EC50 of 2 nM, localizes to EGFR+ tumor xenografts in athymic nude mice and that uptake of the dye in xenografts is significantly reduced when EGFR are blocked by preinjection of excess unlabeled Centyrin. Taken together, these data suggest that CNDCs can be used for intraoperative identification and surgical removal of EGFR-expressing lesions and that Centyrins targeted to other tumor-specific antigens should prove similarly useful in fluorescence guided surgery of cancer. In addition, we demonstrate that the CNDC is detected in the NIR region of the spectrum and can be utilized for fluorescence-guided surgery (FGS). In addition, we propose that with its eventual complete clearance from EGFR-negative tissues and its quantitative retention in the tumor mass for >24 h, a Centyrin-targeted NIR dye should provide excellent tumor contrast when injected at least 6-8 h before initiation of cancer surgery in human patients.
Insights
This study introduces a novel near-infrared fluorescent dye, a Centyrin-near-infrared dye conjugate (CNDC), for improved cancer surgery. CNDCs enable precise visualization and removal of epidermal growth factor receptor-expressing tumors during fluorescence-guided surgery.
Area of Science:
- Biomedical Engineering
- Oncology
- Surgical Technology
Background:
- Near-infrared (NIR) fluorescent dyes enhance cancer surgery by improving visualization of malignant lesions.
- Current dyes include small molecules and antibodies, but low molecular weight protein scaffolds offer superior tumor penetration and antigen-binding versatility.
Purpose of the Study:
- To develop and evaluate a Centyrin-based NIR fluorescent dye conjugate for fluorescence-guided surgery (FGS) targeting epidermal growth factor receptor (EGFR)-expressing tumors.
- To optimize imaging parameters for real-time FGS in preclinical models.
Main Methods:
- A 10 kDa protein scaffold (Centyrin) was conjugated to a NIR fluorescent dye.
- The Centyrin-near-infrared dye conjugate (CNDC) was tested for binding affinity to EGFR-positive cancer cells (EC50 = 2 nM).
- Tumor targeting and imaging were assessed in EGFR-expressing tumor xenografts in athymic nude mice, with and without EGFR blockade.
Main Results:
- CNDC demonstrated selective binding to human EGFR+ cancer cells.
- CNDC localized effectively to EGFR+ tumor xenografts in vivo.
- Pre-blocking EGFR significantly reduced CNDC uptake in xenografts, confirming target specificity.
- The CNDC is detectable in the NIR spectrum and suitable for FGS.
Conclusions:
- CNDC enables intraoperative identification and surgical resection of EGFR-expressing tumors.
- Centyrin scaffolds can be engineered to target various tumor antigens for FGS.
- CNDCs offer excellent tumor contrast for FGS, with potential for clinical application 6-8 hours post-injection.
