Live Cell BH3 Profiling for Single-Cell Drug Response Imaging
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Existing methods for precision medicine in cancer treatment rely on static genetic information and fail to account for the functional complexity of cancer cells, often limited by low cell yields from biopsies and lack the ability to distinguish between cancerous and non-cancerous cells, and do not permit dynamic assessment of pro-apoptotic signaling.
Innovation Solution
A method involving live cell imaging and BH3 peptide-induced mitochondrial outer membrane permeabilization (MOMP) measurement in primary cancer cells, allowing for dynamic BH3 profiling (DBP) to identify therapeutic agents by comparing MOMP in treated and untreated cells, and adjusting BH3 peptide concentrations to optimize measurements.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If core needle biopsies are used to collect patient samples, then the procedure is minimally invasive, but the cell yield is low
Solution Approach 1:
The patent applies preliminary action by performing live cell imaging and BH3 peptide treatment on cells immediately after collection, before they undergo fixation or other processing steps that would reduce viability. This time-sensitive approach maximizes the utility of the limited cells obtained from minimally invasive biopsies.
Solution Approach 2:
The patent changes the parameter of cell state from fixed/dead to live/functional by performing all measurements while cells are still viable. This includes measuring mitochondrial membrane potential, calcium flux, and other dynamic parameters that require living cells, thereby extracting maximum information from the limited sample quantity.
2Device complexity
If static genetic information is used to link cancer biology, then the analysis is simple, but the functional complexity of cancer cells is lost
Solution Approach 1:
The patent transitions from static genetic analysis to dynamic functional assessment by measuring real-time changes in mitochondrial membrane potential, calcium flux, and other cellular parameters in response to BH3 peptide treatment. This captures the functional state and response dynamics of cancer cells, preserving their complexity.
Solution Approach 2:
The patent substitutes mechanical/genetic analysis with functional/biochemical measurement systems. Instead of relying solely on static DNA sequencing, the system uses live cell imaging, electrophysiological measurements, and biochemical assays to directly measure cellular functions and responses.
3Reliability
If prior art methodologies are used, then the procedures are established, but they do not permit determination of pro-apoptotic signaling on a single cell basis
Solution Approach 1:
The patent applies segmentation by measuring pro-apoptotic signaling parameters at the single-cell level rather than averaging across populations. Each cell's mitochondrial membrane potential, calcium flux, and BH3 peptide response are individually quantified, enabling identification of heterogeneous responses within the cancer cell population.
Solution Approach 2:
The patent uses fluorescent dyes and colorimetric indicators that change color or fluorescence intensity in response to specific cellular events (e.g., mitochondrial membrane potential changes, calcium flux). These optical signals provide real-time, single-cell resolution measurements of pro-apoptotic signaling dynamics.
4Quantity of substance
If limited cell quantities are available, then the sample quantity is constrained, but the number of drugs that can be tested is reduced
Solution Approach 1:
The patent makes the limited cell sample universal by performing multiple types of measurements on the same cells: mitochondrial membrane potential, calcium flux, BH3 peptide binding, and functional response assays. This multi-functional approach extracts maximum information from each cell, enabling comprehensive drug screening despite limited quantity.
Solution Approach 2:
The patent maintains continuous measurement of cellular parameters throughout the BH3 peptide treatment and imaging process. By continuously monitoring mitochondrial membrane potential and other dynamic parameters across multiple time points and drug concentrations, the system maximizes the data extracted from each limited cell sample.
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
Enables the prediction of clinical response to drug combinations by identifying putative therapeutic agents through dynamic assessment of pro-apoptotic signaling in individual cancer cells, even with limited sample quantities, and distinguishing between cancerous and non-cancerous cells.
Implementation Method 1
measuring BH3 peptide-induced mitochondrial outer membrane permeabilization (MOMP) in the test cell portion
Implementation Method 2
capturing a series of images of the test cell portion over a time interval by live cell imaging
Data Source
AI summary
Aspects of the application provide methods and compositions for identifying and evaluating putative therapeutic agents for cancer by live cell imaging. Cell samples comprising cancerous cells that have been pre-treated with a test agent are contacted with a BH3 peptide, and samples are imaged by live cell imaging over a time interval. Methods of the application can be used to determine whether a patient is likely to benefit from treatment with a particular test agent.