MYH11/NDE1 Methylation Markers for Direct EPC Identification
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Solution Overview
Problem
Current methods lack specific and robust tools for identifying and quantifying endothelial progenitor cells (EPCs) in complex samples, particularly in whole blood and non-trypsinized tissues, due to the absence of unique markers that differentiate them from other cell types.
Innovation Solution
Analyze the methylation status of specific CpG positions in the MYH11/NDE1 gene region, specifically demethylation patterns, to identify and quantify EPCs without the need for cell purification or enrichment, using bisulfite convertibility and PCR-based methods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional cell surface marker methods are used to identify EPCs, then identification can be performed, but the methods lack specificity and cannot reliably distinguish EPCs from other cell types in complex samples
Solution Approach 1:
The invention changes the identification parameter from cell surface markers (proteins) to DNA methylation status (epigenetic modification). This parameter change enables specific identification of EPCs through the unique demethylated state of the MYH11/NDE1 gene region, which is not present in other cell types, thereby resolving the contradiction between identification capability and discrimination accuracy
Solution Approach 2:
The invention introduces an intermediary approach by using epigenetic modifications (DNA methylation patterns) as a mediator between cell identity and detection. The methylation status of specific CpG positions in the MYH11/NDE1 region serves as an intermediary marker that specifically identifies EPCs without cross-reactivity with other cell types, solving the specificity problem
2Quantity of substance
If cell purification and enrichment steps are performed before identification, then cell concentration is improved, but the process complexity and time consumption increase
Solution Approach 1:
The invention extracts and analyzes only the specific epigenetic signature (methylation status of MYH11/NDE1 region) directly from the complex sample without requiring prior cell separation. This extraction of the diagnostic marker from the complex biological matrix eliminates the need for purification steps while maintaining identification accuracy
Solution Approach 2:
The invention segments the identification process from the cell preparation process by using a molecular marker that can be detected directly in crude samples. The epigenetic analysis is segmented as an independent detection step that does not require upstream cell enrichment, simplifying the overall workflow
3Measurement precision
If cell purification steps are performed, then cell identification accuracy may be improved, but the processing time and operational complexity increase
Solution Approach 1:
The invention performs preliminary action by targeting a molecular marker (DNA methylation pattern) that is inherently present and stable in EPCs without requiring cell activation, enrichment, or differentiation steps. This preliminary targeting of the epigenetic marker enables direct identification from fresh samples, eliminating time-consuming processing steps
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
Provides a reliable and robust method for distinguishing EPCs from other cells, enabling accurate identification and quantification in complex samples, suitable for clinical applications and disease diagnostics.
Implementation Method 1
analyzing epigenetic modifications/ properties of (including the methylation status) of at least one CpG position in the mammalian gene region for muscle myosin heavy chain 11 (MYH11) and nuclear distribution protein nudE homolog 1 (NDE1)
Data Source
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AI summary
The present invention relates to a method, in particular an in vitro method, for identifying endothelial progenitor cells (EPCs), comprising analyzing epigenetic modifications/ properties of (including the methylation status) of at least one CpG position in the mammalian gene region for muscle myosin heavy chain 11 (MYH11) and nuclear distribution protein nudE homolog 1 (NDE1), wherein a demethylation or lack of methylation of said gene regions is indicative for an EPC, when compared to a non-EPC. The analysis according to the invention can identify EPCs on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood, non-blood or immune cells. The present invention furthermore provides an improved method for quantifying EPCs, in particular in complex samples. The method can be performed without a step of purifying and/or enriching cells, preferably in whole blood and/or non-trypsinized tissue.