Global Health Technologies
Wenting Gao
PhD Candidate
Cornell University
Ithaca, New York, United States
Iftak Hussain
Post-Doc
Cornell University, United States
David Erickson
Professor
Cornell University
Ithaca, New York, United States
The rise of breast cancer incidence in low- and middle-income countries (LMICs) is alarming. As per WHO's estimates, by 2020, 70% of 16 million new global cases per annum will occur in these regions. In high-income countries (HICs), the surge is primarily driven by hormone receptor (HR)-rich cancers, with the prevalence among women aged 40 to 69 increasing from 75.4% to 77.5% between 1992 and 1998. In stark contrast, HR-poor tumors are postulated to fuel the growing incidence in LMICs.
As breast cancer rates escalate in LMICs, it's crucial to ascertain the dominant HR tumor types among the new cases and their contributing factors. Nevertheless, the required studies are significantly impeded by the prohibitive costs and scarce availability of molecular subtyping of breast cancer in LMICs.
Our project confronts this pressing issue by developing a cost-effective, innovative technology for rapid molecular subtyping of breast cancers in LMICs. This ground-breaking technology facilitates a transition to biology-driven patient management, ensuring optimal patient care, effective use of limited health resources, and ultimately, improved cancer treatment outcomes.
Our project aims to employ quantitative, multiplexed Lateral-flow Immunoassay (LIA) technology to establish a kit for rapid quantification of ER, PR, and HER2 levels in Fine Needle Aspirant (FNA) from breast cancers (Figure 1). We will corroborate the results with Immunohistochemistry (IHC) and conduct a proof-of-concept validation study in Nigeria. The proposed workflow involves the introduction of a cell sample obtained from patient or tumor cell culture into a pre-filled micro-centrifuge tube containing a cell lysis mixture (Steps 1-2). The sample is then heated to 65°C, promoting cell lysis and the release of sex hormone receptors (Step 3). Once lysis is achieved, a portion of the sample is pipetted onto a 4-plex lateral flow test strip, featuring color-coded lines for each receptor and a control (Figure 2, Steps 4-5). An at-home portable reader system will analyze the test strip, delivering quantitative results via handheld mobile devices (Step 6).
Our technology builds upon traditional lateral flow diagnostic technology, chosen for its adaptability and compliance with the World Health Organization’s ASSURED criteria for diagnostic tests. Unlike most commercially available tests that singleplex (test for a single agent), our user-friendly Point-of-Care Breast Cancer Reader System requires minimal training, stands out with its multiplexing capability, providing unbiased detection of causative agents in a single test.
Our team successfully developed individual tests for ER, PR, and HER2, leveraging the quantitative multiplexed LIA technology. We performed a serious of experiments using spiked buffers in the range of 0 - 1000 pM (spanning the clinical range) to determine limit of detection, limit of quantification, range of quantification, accuracy and CV for each of the three biomarkers. Each assay displayed robustness, high sensitivity, and specificity, with minimal cross-reactivity between each antibodies. Subsequently, these assays were combined into a single, multiplexed system, capable of simultaneous detection of all three biomarkers. The accuracy of this novel approach will be validated using IHC reference standards.
The results confirmed the feasibility of the Point-of-Care Breast Cancer Reader System in delivering rapid, accurate detection and quantification of ER, PR, and HER2 levels in antigen spiked buffers. The integration of these tests into a single multiplexed system stands as a significant leap forward in point-of-care diagnostics. This approach allows for simultaneous evaluation of multiple hormone receptors, potentially improving the accuracy of breast cancer subtyping and fostering personalized treatment strategies. Further, the system's ease of use and quick turnaround time could have substantial implications for breast cancer care, especially in LMICs where diagnostic resources are scarce. Future steps involve validation of this technology using FNA-derived human samples from breast cancer patients to further test the specificity, sensitivity, and robustness of the assay development, advancing towards our ultimate goal of transforming breast cancer care through accessible, innovative diagnostic technology.
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