Fusion Polypeptide for Universal Allogeneic Cell Therapy
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current genome-editing methodologies for producing allogeneic engineered cells for adoptive immunotherapy are hindered by off-target effects, complex manufacturing processes, and inefficiencies such as double-stranded DNA breaks and the need for separate gene-delivery steps, which can lead to reduced yields and increased risks of graft-versus-host disease and host-versus-graft rejection.
Innovation Solution
A single engineered fusion protein is expressed that simultaneously downregulates the cell surface expression of native MHC class I and native TCR molecules by combining a domain capable of downregulating MHC class I with a domain binding to the TCR/CD3 complex, allowing for co-expression with CARs or transgenic TCRs, thereby simplifying the manufacturing process and reducing off-target effects.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If genome-editing methodologies (CRISPR/Cas9, TALENs, zinc-fingers) are used to disrupt TCR or HLA genes, then permanent abrogation of TCR or HLA is achieved, but off-target effects and translocations occur
Solution Approach 1:
The invention extracts and removes the harmful genome-editing step entirely, replacing it with a transduction-based approach where a viral vector delivers a polypeptide that binds to and downregulates TCR/CD3 complex and MHC class I molecules. This eliminates off-target effects and translocations while achieving the desired functional abrogation.
Solution Approach 2:
The invention introduces an intermediary polypeptide (fusion protein) that mediates between the engineered cell and the target molecules (TCR/CD3 complex and MHC class I). This polypeptide binds to the target molecules and prevents their cell surface expression, avoiding direct genetic disruption.
2Manufacturing precision
If separate gene-delivery steps are used for CAR/TCR transduction and genome editing, then specific gene modification is achieved, but manufacturing complexity increases
Solution Approach 1:
The invention merges the transduction of CAR/TCR genes with the delivery of the downregulating polypeptide into a single viral vector system. This allows both functions to be achieved simultaneously in one manufacturing step, reducing complexity while maintaining precision.
Solution Approach 2:
The viral vector system is designed to be universal, capable of simultaneously delivering multiple functions: CAR/TCR gene expression and polypeptide downregulation. This multi-functional approach eliminates the need for separate delivery steps.
3Reliability
If genome editing is performed at two loci (TCR and HLA), then dual downregulation is achieved, but double-stranded DNA breaks and translocations occur
Solution Approach 1:
The invention extracts the harmful DNA break mechanism entirely by replacing genome editing with protein-based downregulation. The polypeptide binds to TCR/CD3 complex and MHC class I molecules, preventing their surface expression without causing DNA breaks or translocations.
Solution Approach 2:
The fusion protein acts as an intermediary that simultaneously targets both TCR/CD3 complex and MHC class I molecules. By binding to these molecules and preventing their cell surface expression, it achieves dual downregulation without direct genetic manipulation.
4Adaptability or versatility
If autologous engineered cells are manufactured for each patient, then patient-specific customization is achieved, but time delay and manufacturing failure increase
Solution Approach 1:
The invention segments the manufacturing process into two independent parts: (1) pre-manufacturing of allogeneic engineered cells with standardized CAR/TCR and downregulating polypeptide, and (2) post-receiving administration to the patient. This allows parallel processing and eliminates time delays associated with patient-specific manufacturing.
Solution Approach 2:
The invention uses allogeneic donor cells as copies that can be manufactured in advance and then administered to patients. These standardized copies eliminate the need for time-consuming patient-specific manufacturing while maintaining therapeutic effectiveness.
5Productivity
If allogeneic engineered cells are used, then economies of scale are achieved, but graft-versus-host disease and host-versus-graft rejection occur
Solution Approach 1:
The invention extracts and removes the harmful recognition mechanisms (TCR and MHC class I) that cause GvHD and HvGR. By using the polypeptide to bind to and downregulate these molecules, the harmful immune recognition is eliminated while preserving the therapeutic function.
Solution Approach 2:
The fusion protein serves as an intermediary that simultaneously blocks both GvHD and HvGR mechanisms. By binding to TCR/CD3 complex and MHC class I molecules, it prevents the harmful immune interactions while allowing the engineered cell to maintain its therapeutic CAR/TCR function.
Data Source
AI summary
The present invention provides a polypeptide comprising: (i) a first domain which is capable of downregulating cell surface expression of an MHC class I molecule; and (ii) a second domain which is capable of binding to a target molecule.