Flow Cytometry DTC Detection in Bone Marrow Aspirates
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for identifying disseminated tumor cells (DTCs) in bone marrow aspirates are insensitive, fail to detect cells that have undergone endothelial to mesenchymal transition (EMT), and do not enable molecular analysis of the identified DTCs.
Innovation Solution
A composition comprising labeled molecules that specifically bind to markers expressed by DTCs, such as HER2, EpCAM, CD49C, EGFR, c-Met, and pan-cytokeratin, is used to detect DTCs in a sample from a subject. The method involves contacting the sample with the labeled molecules and analyzing it using flow cytometry.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If density gradient centrifugation and immunohistochemistry are used to identify DTCs, then the method is simple to operate, but the detection sensitivity is low and cell loss occurs
Solution Approach 1:
The patent replaces the mechanical density gradient centrifugation method with a biological recognition system using antibodies that specifically bind to DTC markers. This substitution enables direct detection of DTCs in bone marrow aspirates without requiring complex centrifugation steps, thereby improving detection sensitivity while maintaining operational simplicity through standardized immunological assays.
Solution Approach 2:
The patent changes the detection parameters by using multiple specific markers (EpCAM, cytokeratins, HER2, EGFR, c-Met) instead of general morphological criteria. This multi-parameter approach allows for more sensitive and specific identification of DTCs, including those that have undergone EMT, while the use of standardized antibody-based assays keeps the procedure operationally simple.
2Measurement precision
If immunohistochemistry is used to detect DTCs, then the procedure is straightforward, but it misses DTCs that have undergone endothelial to mesenchymal transition (EMT)
Solution Approach 1:
The patent employs a universal marker panel that includes EpCAM, cytokeratins, HER2, EGFR, and c-Met, which collectively detect various phenotypes of DTCs including those that have undergone EMT. This multi-functional marker set ensures comprehensive detection coverage across different DTC states while the panel is designed to be practically implementable with standard flow cytometry or immunohistochemistry protocols.
Solution Approach 2:
The patent uses a composite detection approach combining multiple antibody markers that recognize different surface and intracellular components of DTCs. This composite strategy ensures that even if some markers are downregulated due to EMT, other markers remain positive, allowing detection of the full spectrum of DTC phenotypes without requiring overly complex procedures.
3Loss of information
If conventional methods are used to identify DTCs, then the workflow is simple, but molecular analysis of the identified DTCs is not enabled
Solution Approach 1:
The patent performs preliminary enrichment and isolation of DTCs using specific antibody markers before molecular analysis. By pre-concentrating and purifying the DTC population using flow cytometry or magnetic bead separation based on marker expression, the method preserves molecular integrity and enables subsequent genomic, transcriptomic, or proteomic analyses without requiring overly complex workflows.
Solution Approach 2:
The patent uses antibody-marked beads or fluorescently labeled antibodies as intermediaries to isolate DTCs while preserving their molecular content. These intermediary reagents enable the transition from simple detection to complex molecular analysis by providing a gentle, specific isolation method that maintains cell viability and molecular integrity for downstream applications.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
The method effectively detects DTCs in bone marrow aspirates, enabling the identification of breast cancer patients at increased risk of tumor recurrence and allowing for molecular analysis of the isolated DTCs.
Implementation Method 1
at least one labeled molecule that specifically binds to at least one marker expressed by the DTC
Implementation Method 2
analyzing the sample using flow cytometry
Implementation Method 3
at least one labeled molecule that specifically binds to at least one marker expressed by the DTC
Data Source
AI summary
The present invention provides compositions and methods for detecting and isolating disseminated tumor cells (DTCs) from breast cancer in a bone marrow aspirate of a subject. The invention further provides methods of treating breast cancer in the subject.