PDCD1 Methylation Analysis for PD1+ Cell Identification
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for detecting and quantifying PD1+ cells in complex samples, such as blood or tissues, are not robust and often require purification or enrichment steps, limiting their reliability and convenience.
Innovation Solution
A method analyzing the methylation status of specific CpG positions in the PDCD1 gene region using bisulfite convertibility, allowing for the identification and quantification of PD1+ cells without purification, by distinguishing demethylated regions indicative of PD1+ cells from non-PD1+ cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current methods are used to detect and quantify PD1+ cells, then detection can be performed, but the methods are not robust and require purification or enrichment steps
Solution Approach 1:
The invention extracts and utilizes the specific epigenetic signature (methylation pattern) of PD1+ cells as the detection target, rather than detecting the cells themselves through complex purification. By focusing on the unique molecular marker (demethylated CpG sites in PDCD1 gene region), the method eliminates the need for cell purification while maintaining detection reliability
Solution Approach 2:
The invention introduces an intermediary approach by using epigenetic markers (DNA methylation status) as a proxy for cell identification. Instead of directly detecting and purifying PD1+ cells, the method detects their unique epigenetic signature, which serves as an intermediary indicator that is easier to measure in complex samples
2Measurement precision
If purification steps are performed to identify PD1+ cells, then detection accuracy may improve, but the process becomes more complex and less convenient
Solution Approach 1:
The method enables direct analysis of complex samples without requiring external purification assistance. The epigenetic marker approach allows the detection system to automatically distinguish PD1+ cells from other cell types based on their intrinsic methylation patterns, eliminating the need for manual or instrument-based purification steps
3Reliability
If conventional detection methods are used, then PD1+ cells can be identified, but the methods lack robustness in complex samples
Solution Approach 1:
The invention focuses detection on a specific local feature - the methylation status of particular CpG sites within the PDCD1 gene region. By targeting this localized epigenetic signature rather than attempting to characterize entire cells or use broad markers, the method achieves high reliability in complex samples while simplifying the detection approach
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
This approach provides a robust and reliable means to detect and quantify PD1+ cells, enabling their identification in complex samples without purification, and offers a valuable tool for diagnosing autoimmune diseases, cancer, and immune-related conditions by measuring methylation patterns in whole blood or non-trypsinized tissues.
Implementation Method 1
analyzing the methylation status (bisulfite convertibility) of at least one CpG position in the mammalian (e.g. human) gene region for Programmed cell death 1 (PDCD1)
Data Source
Figure 1
Figure 1
Figure 1
AI summary
The present invention relates to a method, in particular an in vitro method, for identifying PD1+ cells, comprising analyzing the methylation status of at least one CpG position in the mammalian gene region for Programmed cell death 1 (PDCD1), wherein a demethylation or lack of methylation of said gene region is indicative for a PD1+ cell, when compared to a non-PD1+ cell. The analyses according to the invention can identify PD1+ cells on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood or immune cells. The present invention furthermore provides an improved method for quantifying PD1+ cells, 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.