HLA-Deleted Glomerular Endothelial Cells for Non-HLA Antibody Detection
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Solution Overview
Problem
Current methods for diagnosing non-HLA antibody-mediated rejection in renal allografts are limited by the inability to effectively identify non-HLA antibodies, leading to challenges in predicting and preventing graft rejection, particularly in patients without circulating HLA antibodies.
Innovation Solution
The development of engineered glomerular endothelial cells with reduced expression of human leukocyte antigens (HLA) using CRISPR-Cas9 technology, allowing for the detection of non-HLA antibodies through a cell-based assay by comparing antibody binding signals with a predetermined reference value.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If conventional diagnostic methods are used to detect non-HLA antibodies, then the diagnostic process is simple, but the detection precision and reliability are insufficient
Solution Approach 1:
The patent uses engineered glomerular endothelial cells as an intermediary target to detect non-HLA antibodies. These cells are modified to express specific non-HLA antigens (such as AT1R, ETAR, LG3, vimentin, FLT3, ICAM4, endoglin, and agrin) that serve as mediators for antibody binding, enabling precise detection of non-HLA antibodies that conventional methods cannot identify
Solution Approach 2:
The patent modifies the expression levels of non-HLA antigens on the surface of glomerular endothelial cells through genetic engineering. By controlling the expression parameters of specific antigens, the assay achieves optimized detection sensitivity and specificity for non-HLA antibodies, resolving the contradiction between detection precision and assay complexity
2Ease of manufacture
If HLA-expressing cells are used for antibody detection, then the cells are easier to maintain, but they cannot effectively detect non-HLA antibodies due to HLA interference
Solution Approach 1:
The patent extracts and removes HLA molecules from the surface of glomerular endothelial cells used in the assay. By eliminating HLA expression from these diagnostic cells, the system prevents false-positive detections and ensures that only non-HLA antibodies are detected, thereby improving detection reliability while maintaining cell ease of manufacture
Solution Approach 2:
The patent creates cells with localized differential properties: the diagnostic glomerular endothelial cells are engineered to lack HLA expression but retain or overexpress specific non-HLA antigens. This local quality differentiation enables selective detection of non-HLA antibodies while maintaining cell viability and ease of cultivation
3Adaptability or versatility
If multiple non-HLA antibody targets are tested, then the detection coverage is improved, but no single test can identify all non-HLA antibodies
Solution Approach 1:
The patent creates a universal diagnostic platform using engineered glomerular endothelial cells that can detect multiple types of non-HLA antibodies simultaneously. The cells are designed to express multiple non-HLA antigens on their surface, enabling a single assay to screen for various non-HLA antibodies (anti-AT1R, anti-ETAR, anti-LG3, anti-vimentin, anti-FLT3, anti-ICAM4, anti-endoglin, and anti-agrin antibodies), thereby achieving broad detection coverage without requiring multiple separate tests
Solution Approach 2:
The patent merges multiple antigen expression functions into a single cell type. The engineered glomerular endothelial cells simultaneously express multiple non-HLA antigens that would otherwise require separate assays, combining multiple detection functions into one unified test system that reduces overall testing complexity
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 enables sensitive and reproducible detection of non-HLA antibodies, improving the prediction and prevention of antibody-mediated rejection, thereby reducing the risk of graft rejection and informing tailored treatment strategies.
Implementation Method 1
The development of engineered glomerular endothelial cells with reduced expression of human leukocyte antigens (HLA) using CRISPR-Cas9 technology
Implementation Method 2
obtaining a quantification of a predetermined signal of non-HLA antibodies bound to said engineered human glomerular endothelial cell
Data Source
AI summary
The invention relates to an engineered glomerular endothelial cell comprising a reduction in expression of a human leukocyte antigen and its use in an in vitro method for determining the likelihood of occurrence of a non-HLA antibody mediated rejection against a renal allograft in an individual in need thereof. The inventors have observed that by suppressing expression of HLA class I and II molecules it was possible to obtain a cell able to bind non-HLA antibodies. The in vitro diagnostic method has been validated on a cohort of patients.