Designer Platelets via CRISPR Editing for Alloantigen Matching
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for diagnosing and treating platelet-related disorders, such as neonatal alloimmune thrombocytopenia, are hindered by the lack of available platelets expressing rare alloantigens, leading to difficulties in antibody detection and transfusion therapy due to the presence of HLA antigens that can cause cross-reactivity.
Innovation Solution
The development of 'designer platelets' using CRISPR/Cas9 gene editing techniques to express specific human platelet alloantigens in induced pluripotent stem cells, which can be differentiated into megakaryocytes and platelets, allowing for the creation of platelets lacking HLA class I antigens to simplify diagnostic testing and provide antigen-matched transfusions.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional platelet transfusion methods are used, then patients can receive platelets from donors, but HLA antigens on platelets cause cross-reactivity and complicate antibody detection
Solution Approach 1:
The patent removes HLA class I antigens from platelets through CRISPR/Cas9 gene editing by targeting the beta-2 microglobulin gene, which is essential for HLA class I antigen expression. This extraction of the harmful HLA antigens eliminates cross-reactivity issues while preserving platelet function and alloantigen expression for accurate antibody detection
Solution Approach 2:
The patent applies local quality modification by selectively eliminating only HLA class I antigens from platelets while preserving other important platelet surface antigens. This localized genetic modification allows the platelets to maintain their diagnostic value for detecting patient antibodies against platelet alloantigens without the interfering presence of HLA antigens
2Reliability
If platelets expressing rare alloantigens are needed for diagnostic testing, then antibody detection can be performed, but available platelet sources are limited and difficult to obtain
Solution Approach 1:
The patent performs preliminary action by pre-engineering platelets with specific rare alloantigens through CRISPR/Cas9 gene editing before they are needed for diagnostic testing. By creating a bank of designer platelets with known alloantigen profiles in advance, the system eliminates the difficulty of sourcing rare platelet types when diagnostic needs arise
Solution Approach 2:
The patent uses induced pluripotent stem cells as a renewable source to produce copies of platelets with specific alloantigen profiles. This copying approach allows unlimited production of platelets expressing rare alloantigens that would otherwise be difficult to obtain from natural donor pools
3Adaptability or versatility
If CRISPR/Cas9 gene editing is applied to induce pluripotent stem cells to create designer platelets, then platelets with specific alloantigens can be produced, but the process complexity increases
Solution Approach 1:
The patent employs universal CRISPR/Cas9 gene editing protocols that can be applied to create platelets with any desired alloantigen profile using the same basic methodology. This universal approach, combined with induced pluripotent stem cell technology that can differentiate into multiple blood cell types, provides a versatile platform for producing designer platelets with various alloantigens without requiring separate specialized processes for each antigen type
Data Source
AI summary
Methods of creating cells expressing specific platelet alloantigens by combining gene editing techniques and cell culture differentiation or expansion techniques employing pluripotent cells, including the steps of transfecting pluripotent cells with a plasmid encoding one or more guide RNAs targeting within a platelet alloantigen target locus and a nuclease in the presence of an HDR repair oligo and culturing the resulting cells to expand their numbers or to create a differentiated cell type of interest.