Recombinant Adipose-Derived Stem Cell for Hemophilia B Therapy
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Solution Overview
Problem
Current gene therapy methods for hemophilia B, such as in vivo and ex vivo pathways, face challenges including high immune response, viral inactivation, and complex, costly procedures, leading to inefficient expression of coagulation factor IX and prolonged treatment cycles.
Innovation Solution
Transfecting an adenovirus carrying a human factor IX gene into adipose-derived stem cells, which are then transfected at 80% confluency under specific conditions, to obtain recombinant cells that continuously and efficiently express the hFIX protein.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If in vivo gene therapy pathway is used to directly inject vector carrying FIX gene into patient, then operation convenience and treatment speed are improved, but immune response and safety risk increase
Solution Approach 1:
The patent introduces adipose-derived stem cells as an intermediary carrier to deliver the FIX gene. Instead of directly injecting the virus into the patient (in vivo), the virus-transduced ADSCs are first prepared ex vivo and then infused into the patient. This intermediary step reduces direct immune confrontation with the viral vector while maintaining gene delivery effectiveness.
Solution Approach 2:
The patent performs gene transduction and cell preparation in advance (ex vivo) before patient infusion. The ADSCs are pre-transduced with the FIX gene in a controlled laboratory setting, allowing for optimization of transduction efficiency and cell quality before clinical application, thereby reducing the need for high-dose viral vectors during actual treatment.
2Reliability
If ex vivo gene therapy pathway is used to transfer gene into somatic cell during in vitro culture, then safety and controllability are improved, but operation complexity and treatment cycle increase
Solution Approach 1:
The patent uses adipose-derived stem cells which possess multiple functions: they serve as both the target cell for gene delivery and as a functional cell type that can secrete FIX protein. Additionally, ADSCs have self-renewal capability and can be easily harvested from patients, combining safety, efficacy, and operational simplicity in a single cell type.
Solution Approach 2:
The patent optimizes specific parameters including transduction efficiency (achieving high levels of FIX gene expression in ADSCs), cell passage number (using early passage cells for better transduction efficiency), and viral titer (controlling the multiplicity of infection) to balance safety and operational simplicity.
3Productivity
If high level expression of FIX factor is achieved in hemophilia animals, then therapeutic effect is improved, but expression quantity decreases over time due to immunogenic response
Solution Approach 1:
The patent uses ADSCs as an intermediary that secretes FIX protein rather than directly expressing it from transduced cells. The ADSCs serve as a protected factory, shielding the immunogenic FIX protein from direct immune recognition while maintaining continuous secretion, thereby extending the duration of therapeutic effect.
Solution Approach 2:
The patent utilizes the endogenous secretory capacity of adipose-derived stem cells to continuously produce and secrete FIX protein. The ADSCs' natural self-renewal and proliferation capabilities enable long-term, sustained production of therapeutic FIX without requiring repeated administrations, making the system self-sustaining.
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
The recombinant adipose-derived stem cells effectively express and secrete the hFIX protein, potentially converting severe hemophilia B to a milder form by achieving stable FIX activity levels, reducing immune response risks and treatment complexity.
Implementation Method 1
transfecting an adenovirus carrying a human factor IX gene into adipose-derived stem cells
Data Source
AI summary
The present invention provides a recombinant adipose-derived stem cell and a recombinant method thereof, and belongs to the technical field of genetic engineering, where an adenovirus carrying an hFIX gene is transfected into an adipose-derived stem cell to obtain the recombinant adipose-derived stem cell. In the present invention, an adenovirus carrying an hFIX gene is transfected into an adipose-derived stem cell, and the recombinant adipose-derived stem cell obtained after the transfection can express an hFIX protein.